RAD51 inhibitors

ABSTRACT

This application is directed to inhibitors of RAD51 represented by the following structural formula, 
                         
and methods for their use, such as to treat cancer, autoimmune diseases, immune deficiencies, or neurodegenerative diseases.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/556,763, filed on Sep. 11, 2017; and U.S. Provisional Application No.62/711,959, filed on Jul. 30, 2018. The entire teachings of theaforementioned applications are incorporated herein by reference.

FIELD OF THE INVENTION

This application is directed to inhibitors of RAD51, and methods fortheir use, such as to treat conditions including cancers, autoimmunediseases, immune deficiencies, and neurodegenerative diseases.

BACKGROUND OF THE INVENTION

RAD51 is a eukaryote gene. The protein encoded by this gene is a memberof the RAD51 protein family which assists in repair of DNA double strandbreaks. RAD51 family members are homologous to the bacterial RecA,Archaeal RadA and yeast RAD51. The protein is highly conserved in mosteukaryotes, from yeast to humans. In humans, RAD51 is a 339-amino acidprotein that plays a major role in homologous recombination of DNAduring double strand break (DSB) repair. RAD51 catalyzes strand transferbetween a broken sequence and its undamaged homologue to allowre-synthesis of the damaged region (see homologous recombinationmodels).

Studies have demonstrated a sensitization to certain DNA damagingtherapies associated with cellular defects in proteins that promote HRDNA repair. This sensitization is particularly dramatic for DNAcross-linking chemotherapeutic drugs (30-100 times) and ionizingradiation (3-5 times) (Godthelp et al., “Mammalian Rad51C contributes toDNA cross-link resistance, sister chromatid cohesion and genomicstability,” Nucleic Acids Res., 30:2172-2182, 2002; Tebbs et al.,“Correction of chromosomal instability and sensitivity to diversemutagens by a cloned cDNA of the XRCC3 DNA repair gene,” Proc. Natl.Acad. Sci. USA, 92:6354-6358, 1995; Takata et al., “Chromosomeinstability and defective recombinational repair in knockout mutants ofthe five Rad51 paralogs,” Mol. Cell Biol., 21:2858-2866, 2001; Liu etal., “XRCC2 and XRCC3, new human Rad51-family members, promotechromosome stability and protect against DNA cross-links and otherdamages,” Mol. Cell, 1:783-793, 1998).

Several groups have recently demonstrated that HR can be partiallyinhibited in order to sensitize cells to DNA damaging therapies.Inhibition of XRCC3 (a RAD51 paralog protein), has been demonstratedusing a synthetic peptide corresponding to another paralog protein. Thispeptide sensitized Chinese Hamster Ovary (CHO) cells to cisplatin andinhibited the formation of sub-nuclear RAD51 foci in response to DNAdamage (Connell et al., Cancer Res., 64:3002-3005, 2004). Otherresearchers have inhibited the expression of the RAD51 protein itself(Russell et al., Cancer Res., 63:7377-7383, 2003; Hansen et al., Int. J.Cancer, 105:472-479, 2003; Ohnishi et al., Biochem. Biophys. Res.Commun., 245:319-324, 1998; Ito et al., J. Gene Med., 7(8):1044-1052,2005; Collins et al., Nucleic Acids Res., 29:1534-1538, 2001) or blockedits function by over-expressing a dominant negative BRC peptide fragmentderived from BRCA2 (Chen et al., J. Biol. Chem., 274:32931-32935, 1999).In view of the connection between increased sensitivity to certain DNAdamaging therapies and cellular defects in HR DNA repair-relatedproteins, there is a need for additional compounds that inhibit RAD51.

SUMMARY OF THE INVENTION

Applicant has now discovered novel compounds which are effectiveinhibitors of RAD51 (see Examples 1-75). The RAD51 inhibitors of thepresent invention inhibit homologous recombination by altering thenucleocytoplasmic distribution of RAD51 following DNA damage induction.The RAD51 inhibitors of the present invention reduce the repair ofAID-induced DNA double strand breaks, leading to AID-dependentcytotoxicity in both normal and malignant B-lymphocytes. Certain ofthese RAD51 inhibitors have superior cell permeability as measured inCaco-2 cells (see Example 76). Among the RAD51 inhibitors with good cellpermeability, several have superior metabolic stability (as measured bya liver microsome assay, see Example 77) and exposure, including oralexposure (see Example 79).

The present invention provides a compound represented by StructuralFormula I:

or a pharmaceutically acceptable salt thereof. The definition of eachvariable is provided below.

The present invention also provides a pharmaceutical compositioncomprising a compound as described herein or a pharmaceuticallyacceptable salt thereof and a pharmaceutically acceptable carrier ordiluent.

The present invention further provides a method of treating a cancer, anautoimmune disease, an immune deficiency, or a neurodegenerativedisease. The method comprises administering to a subject in need thereofan effective amount of a compound of disclosed herein or apharmaceutically acceptable salt thereof or a pharmaceutical compositiondisclosed herein.

Also provided is the use of a compound disclosed herein, or apharmaceutically acceptable salt thereof, or a pharmaceuticalcompositions disclosed herein for the preparation of a medicament forthe treatment of a cancer, an autoimmune disease, an immune deficiency,or a neurodegenerative disease.

Also provided is a compound disclosed herein, or a pharmaceuticallyacceptable salt thereof, or a pharmaceutical composition disclosedherein for use in treating a cancer, an autoimmune disease, an immunedeficiency, or a neurodegenerative disease.

Although Applicant does not wish to be bound by any mechanism, it isbelieved that the compounds of the invention inhibit RAD51 by binding toMDC1 and causing reduced ability to form active complexes of RAD51.

DETAILED DESCRIPTION

In a first embodiment, the invention provides a compound represented byStructural Formula I:

or a pharmaceutically acceptable salt thereof, wherein:

the thiazole ring is optionally substituted with —F or —Cl;

Cy is —(C₃-C₇)cycloalkyl, bridged (C₆-C₁₂) cycloalkyl, or a 4-12membered heterocyclic ring, each of which is optionally substituted withone or more groups selected from the group consisting of halogen, —OH,(C₁-C₄)alkyl, and (C₁-C₄)alkoxy;

when X⁵ is connected with a nitrogen ring atom of Cy, X⁵ is absent;

when X⁵ is connected with a carbon ring atom of Cy, X⁵ is NR^(a) or O;

X⁶ is NR^(a) or O;

R¹ is (C₁-C₅)alkyl;

R³ is (C₁-C₅)alkyl, —CH₂-phenyl, —(C₃-C₇)cycloalkyl,—CH₂—(C₃-C₇)cycloalkyl, —CH₂-monocyclic 3-7 membered heterocyclic ring,or monocyclic 3-7 membered heterocyclic ring, wherein the (C₁-C₅)alkyl,—(C₃-C₇)cycloalkyl, phenyl or monocyclic 3-7 membered heterocyclic ringrepresented by R³ or in the group represented by R³ is optionallysubstituted with one or more groups selected from the group consistingof halogen, —OH, (C₁-C₄)alkyl, halomethyl, halomethoxy, —CN, and(C₁-C₄)alkoxy;

R² is —NR^(a)C(O)O(C₁-C₄)alkyl; —NR^(a)C(O)NR^(a)(C₁-C₄)alkyl;—NR^(a)C(O)O(C₂-C₄)alkenyl;

—NR^(a)C(O)NR^(a)(C₂-C₄)alkenyl; —NR^(a)C(O)O—(C₃-C₆)cycloalkyl;—NR^(a)C(O)NR^(a)—(C₃-C₇)cycloalkyl; —NR^(a)C(O)O-phenyl;—NR^(a)C(O)NR^(a)-phenyl; —NR^(a)C(O)O-monocyclic 3-7 memberedheterocyclic ring; —NR^(a)C(O)NR^(a)-monocyclic 3-7 memberedheterocyclic ring; —NR^(a)C(O)O-monocyclic 5-6 membered heteroaromaticring; —NR^(a)C(O)NR^(a)-monocyclic 5-6 membered heteroaromatic ring;

wherein the (C₁-C₄)alkyl and the (C₂-C₄)alkenyl in the group representedby R² are each optionally and independently substituted with one or moregroups selected from the group consisting of halogen, N₃, —OR^(a),—NR^(a)R^(a), —(C₃-C₆)cycloalkyl, phenyl, a monocyclic 3-7 memberedheterocyclic ring, and a monocyclic 5-6 membered heteroaromatic ring;

wherein the (C₃-C₇)cycloalkyl in the group represented by R² isoptionally substituted with one or more groups selected from the groupconsisting of halogen, —CH₃, ═O, —OR^(a) and —NR^(a)R^(a);

wherein the phenyl in the group represented by R² is optionallysubstituted with one or more groups selected from the group consistingof halogen, —CH₃, halomethyl, halomethoxy,

—CN, —OR^(a), and —N₃;

wherein the heterocyclic ring in the group represented by R² isoptionally substituted with one or more groups selected from the groupconsisting of ═O, halogen, —OR^(a), —CH₃, halomethyl, and halomethoxy;

wherein the heteroaromatic ring in the group represented by R² isoptionally substituted with one or more groups selected from the groupconsisting of halogen, —CN, —CH₃, halomethyl, halomethoxy, —OR^(a) and—NR^(a)R^(a); and

each R^(a) is independently —H or —CH₃.

In a second embodiment, the invention provides a compound represented byStructural Formula II:

or a pharmaceutically acceptable salt thereof, wherein

the thiazole ring is optionally substituted with —F or —Cl;

Cy is cyclohexyl or a 6-membered monocyclic heterocyclic ring;

X⁵ and X⁶ are each independently NR^(a) or O;

R¹ is (C₁-C₅)alkyl;

R³ is (C₁-C₅)alkyl or monocyclic 3-7-membered heterocyclic ring;

R² is —NR^(a)C(O)O(C₁-C₄)alkyl; —NR^(a)C(O)NR^(a)(C₁-C₄)alkyl;—NR^(a)C(O)O(C₂-C₄)alkenyl;

—NR^(a)C(O)NR^(a)(C₂-C₄)alkenyl; —NR^(a)C(O)—O(C₃-C₆)cycloalkyl;—NR^(a)C(O)NR^(a)—(C₃-C₆)cycloalkyl; —NR^(a)C(O)O-phenyl;—NR^(a)C(O)NR^(a)-phenyl; —NR^(a)C(O)O-monocyclic 3-7 memberedheterocyclic ring; —NR^(a)C(O)NR^(a)-monocyclic 3-7 memberedheterocyclic ring; —NR^(a)C(O)O-monocyclic 5-6 membered heteroaromaticring; —NR^(a)C(O)NR^(a)-monocyclic 5-6 membered heteroaromatic ring;

wherein the (C₁-C₄)alkyl and the (C₂-C₄)alkenyl in the group representedby R² are each optionally and independently substituted with one or morehalogen, N₃, —OR^(a), —NR^(a)R^(a), —(C₃-C₆)cycloalkyl, phenyl,monocyclic 3-7-membered heterocyclic ring, or monocyclic 5-6-memberedheteroaromatic ring;

wherein the —(C₃-C₆)cycloalkyl in the group represented by R² isoptionally substituted with one or more halogen, —CH₃, —OR^(a) or—NR^(a)R^(a);

wherein the phenyl in the group represented by R² is optionallysubstituted with one or more halogen, —CH₃, halomethyl, halomethoxy,—OR^(a), or —N₃;

wherein the heterocyclic ring in the group represented by R² isoptionally substituted with one or more ═O, halogen, —CH₃, halomethyl,or halomethoxy;

wherein the heteroaromatic ring in the group represented by R² isoptionally substituted with one or more halogen, —CH₃, halomethyl,halomethoxy, —OR^(a) or —NR^(a)R^(a); and

each R^(a) is independently —H or —CH₃.

In a third embodiment, the invention provides a compound according toStructural Formula I, or a pharmaceutically acceptable salt thereof,wherein Cy is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl; azetidinyl, azepanyl, diazaspiro[4.4]nonyl,diazaspiro[3.5]nonyl, diazepanyl, dihydroimidazole, dihydrofuranyl,dihydropyranyl, dihydropyridinyl, dihydropyrimidinyl, dihydrothienyl,dihydrothiophenyl, dihydrothiopyranyl, hexahydropyridazinyl,hexahydropyrimidinyl, hydantoinyl, indolinyl, isoindolinyl, morpholinyl,oxiranyl, oxetanyl, piperidinyl, piperazinyl, pyrrolidinonyl,pyrrolidinyl, tetrahydrofuranyl, tetrahydroimidazole, tetrahydroindolyl,tetrahydropyranyl, tetrahydrothienyl, tetrahydropyridinyl,tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl,thiomorpholinyl, tropanyl, valerolactamyl; bicyclo[2.1.1]hexyl,bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, bicyclo[3.2.1]octyl,bicyclo[4.3.1]decyl, bicyclo[3.3.1]nonyl, bornyl, bornenyl, norbornyl,norbornenyl, 6,6-dimethylbicyclo [3.1.1]heptyl, tricyclobutyl,adamantly; azanorbornyl, quinuclidinyl, isoquinuclidinyl, tropanyl,azabicyclo[2.2.1]heptanyl, 2-azabicyclo[3.2.1]octanyl,azabicyclo[3.2.1]octanyl, azabicyclo[3.2.2]nonanyl,azabicyclo[3.3.0]nonanyl, azabicyclo [3.3.1]nonanyl,diazabicyclo[2.2.1]heptanyl, diazabicyclo[3.2.1]octanyl,octahydropyrrolo[3,4-b]pyrrolyl, octahydropyrrolo[3,4-c]pyrrolyl; andthe remaining variables are as defined in the first embodiment.

In a fourth embodiment, the invention provides a compound according toStructural Formula I or II, or a pharmaceutically acceptable saltthereof, wherein Cy is cyclohexyl, morpholinyl, thiomorpholinyl,piperidinyl, piperazinyl, hexahydropyridazinyl, hexahydropyrimidinyl,valerolactamyl, dihydropyranyl, dihydropyridinyl, dihydropyrimidinyl,dihydrothiopyranyl, tetrahydropyranyl, tetrahydropyridinyl,tetrahydropyrimidinyl, or tetrahydrothiopyranyl; and the remainingvariables are as defined in the first, second, and/or third embodiments.

In a fifth embodiment, the invention provides a compound represented byStructural Formula III,

or a pharmaceutically acceptable salt thereof, wherein:

X⁷ is NH or O;

R⁴ is (C₁-C₄)alkyl, (C₃-C₆)cycloalkyl, or a monocyclic 3-7 memberedheterocyclic ring;

wherein the (C₁-C₄)alkyl represented by R⁴ is optionally substitutedwith one or more groups selected from the group consisting of halogen,N₃, —OR^(a), —NR^(a)R^(a), —(C₃-C₆)cycloalkyl, phenyl, a monocyclic 3-7membered heterocyclic ring, and a monocyclic 5-6 membered heteroaromaticring,

wherein the (C₃-C₆)cycloalkyl or the monocyclic 3-7 memberedheterocyclic ring represented by R⁴, the (C₃-C₆)cycloalkyl or themonocyclic 3-7 membered heterocyclic ring in the group represented by R⁴is optionally substituted with one or more groups selected from thegroup consisting of halogen, —OR^(a), ═O, and —CH₃,

wherein the phenyl in the group represented by R⁴ is optionallysubstituted with one or more groups selected from the group consistingof halogen, —CH₃, halomethyl, halomethoxy,

—OR^(a), and —N₃;

wherein the heteroaromatic ring in the group represented by R⁴ isoptionally substituted with one or more groups selected from the groupconsisting of halogen and —CH₃; and the remaining variables are asdefined in the first, second, third, and/or fourth embodiments.

In a sixth embodiment, the invention provides a compound according toStructural Formula III, or a pharmaceutically acceptable salt thereof,wherein X⁷ is NH or O; R³ is (C₁-C₅)alkyl; and R⁴ is (C₁-C₄)alkylwherein the (C₁-C₄)alkyl represented by R⁴ is optionally substitutedwith one or more halogen, —OR^(a), —NR^(a)R^(a), —(C₃-C₆)cycloalkyl,phenyl (optionally substituted by one or more halogen, —CH₃, halomethyl,halomethoxy, OR^(a) or N₃), monocyclic 3-7-membered heterocyclic ring(optionally substituted by ═O, halogen or —CH₃), or monocyclic5-6-membered heteroaromatic ring (optionally substituted by halogen or—CH₃); and the remaining variables are as defined in the first, second,third, fourth and/or fifth embodiments.

In a seventh embodiment, the invention provides a compound representedby Structural Formula IV,

or a pharmaceutically acceptable salt thereof; and the variables are asdefined in the first, second, third, fourth, fifth and/or sixthembodiments.

In an eighth embodiment, the invention provides a compound representedby Structural Formula V,

or a pharmaceutically acceptable salt thereof; and the variables are asdefined in the first, second, third, fourth, fifth and/or sixthembodiments.

In a ninth embodiment, the invention provides a compound represented byStructural Formula VI:

or a pharmaceutically acceptable salt thereof; and the variables are asdefined in the first, second, third, fourth, fifth and/or sixthembodiments.

In a tenth embodiment, the invention provides a compound represented byStructural Formula VII:

or a pharmaceutically acceptable salt thereof; and the variables are asdefined in the first, second, third, fourth, fifth and/or sixthembodiments.

In an eleventh embodiment, the invention provides a compound representedby Structural Formula VIII:

or a pharmaceutically acceptable salt thereof; and the variables are asdefined in the first, second, third, fourth, fifth and/or sixthembodiments.

In a twelfth embodiment, the invention provides a compound representedby Structural Formula IX:

or a pharmaceutically acceptable salt thereof; and the variables are asdefined in the first, second, third, fourth, fifth and/or sixthembodiments.

In a thirteenth embodiment, the invention provides a compound accordingto Structural Formula I, II, or III, or a pharmaceutically acceptablesalt thereof, wherein Cy is azetidinyl or pyrrolidinyl, and the nitrogenring atom is connected with the thiazole ring; and the remainingvariables are as defined in the first, second, third, fourth, fifthand/or sixth embodiments.

In a fourteenth embodiment, the invention provides a compound accordingto Structural Formula I, II, or III, or a pharmaceutically acceptablesalt thereof, wherein Cy is 1,7-diazaspiro[4.4]nonyl,2,7-diazaspiro[4.4]nonyl, 2,7-diazaspiro[3.5]nonyl, 1,4-diazepanyl,2,5-diazabicyclo[2.2.1]heptanyl, 3,8-diazabicyclo[3.2.1]octanyl,octahydropyrrolo[3,4-b]pyrrolyl, or octahydropyrrolo[3,4-c]pyrrolyl, andthe two nitrogen ring atoms are connected with the thiazole ring and the—X⁵C(O)X⁶R³ moiety, respectively; and the remaining variables are asdefined in the first, second, third, fourth, fifth and/or sixthembodiments.

In a fifteenth embodiment, the invention provides a compound accordingto Structural Formula III, IV, V, VI, VII, VIII, or IX, or apharmaceutically acceptable salt thereof, wherein R⁴ is —(C₁-C₃)alkyl,(C₃-C₆)cycloalkyl, or a monocyclic 3-7 membered heterocyclic ring,wherein the —(C₁-C₃)alkyl is optionally substituted with (i) phenyloptionally substituted by one or more halogen or —CH₃; (ii) a monocyclic5-6 membered heteroaromatic ring optionally substituted by one or morehalogen or —CH₃; or (iii) a monocyclic 3-7 membered heterocyclic ringoptionally substituted by one or more groups selected from the groupconsisting of halogen and —CH₃; and the remaining variables are asdefined in the first, second, third, fourth, fifth, seventh, eighth,ninth, tenth, eleventh, twelfth, thirteenth, and/or fourteenthembodiments.

In a sixteenth embodiment, the invention provides a compound accordingto Structural Formula III, IV, V, VI, VII, VIII, or IX, or apharmaceutically acceptable salt thereof, wherein R⁴ is —(C₁-C₃)alkyl,—CHR^(a)-phenyl, —CHR^(a)-5-6 membered heteraromatic ring, or—CHR^(a)-3-7 membered monocyclic heterocyclic ring, wherein the phenyl,5-6 membered heteraromatic ring or 3-7 membered monocyclic heterocyclicring in the group represented by R⁴ is optionally substituted one ormore groups selected from the group consisting of halogen and —CH₃; andthe remaining variables are as defined in the first, second, third,fourth, fifth, seventh, eighth, ninth, tenth, eleventh, twelfth,thirteenth, and/or fourteenth embodiments.

In a seventeenth embodiment, the invention provides a compound accordingto Structural Formula III, IV, V, VI, VII, VIII, or IX, or apharmaceutically acceptable salt thereof, wherein R⁴ is —(C₁-C₃)alkyl,optionally substituted with (i) phenyl optionally substituted by one ormore halogen, —CH₃, halomethyl, halomethoxy, OR^(a), or N₃; (ii) amonocyclic 5-6-membered heteroaromatic ring optionally substituted byone or more halogen or —CH₃; or (iii) a monocyclic 3-7-memberedheterocyclic ring optionally substituted by one or more ═O or —CH₃; andthe remaining variables are as defined in the first, second, third,fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth,thirteenth, and/or fourteenth embodiments.

In an eighteenth embodiment, the invention provides a compound accordingto Structural Formula III, IV, V, VI, VII, VIII, or IX, or apharmaceutically acceptable salt thereof, wherein R⁴ is —(C₁-C₃)alkyl,optionally substituted with (i) phenyl optionally substituted by one ormore halogen, —CH₃, halomethyl, halomethoxy, OR^(a), or N₃; (ii) amonocyclic 5-6-membered heteroaromatic ring optionally substituted byone or more halogen or —CH₃; or (iii) a monocyclic 3-7-memberedheterocyclic ring optionally substituted by one or more ═O or —CH₃; andthe remaining variables are as defined in the first, second, third,fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth,thirteenth, fourteenth, and/or seventeenth embodiments.

In a nineteenth embodiment, the invention provides a compound accordingto Structural Formula I, III, IV, V, VI, VII, VIII, or IX, or apharmaceutically acceptable salt thereof, wherein R³ is (C₁-C₄)alkyl,—(C₄-C₆)cycloalkyl, —CH₂-phenyl, —CH₂-monocyclic 4-6 memberedheterocyclic ring, or monocyclic 4-6 membered heterocyclic ring, whereinthe phenyl or monocyclic 4-6 membered heterocyclic ring represented byR³ or in the group represented by R³ is optionally substituted with oneor more groups selected from the group consisting of halogen,

—OR^(a), and —CH₃; and the remaining variables are as defined in thefirst, third, fourth, fifth, seventh, eighth, ninth, tenth, eleventh,twelfth, thirteenth, fourteenth, fifteenth, sixteenth, seventeenth,and/or eighteenth embodiments.

In a twentieth embodiment, the invention provides a compound representedby Structural Formula X:

or a pharmaceutically acceptable salt thereof; and the variables are asdefined in the first, second, third, fourth, fifth, sixth, seventh,fifteenth, sixteenth, seventeenth, eighteenth, and/or nineteenthembodiments.

In a twenty first embodiment, the invention provides a compoundrepresented by Structural Formula XI:

or a pharmaceutically acceptable salt thereof; and the variables are asdefined in the first, second, third, fourth, fifth, sixth, seventh,fifteenth, sixteenth, seventeenth, eighteenth, and/or nineteenthembodiments.

In a twenty second embodiment, the invention provides a compoundrepresented by Structural Formula XII:

or a pharmaceutically acceptable salt thereof; and the variables are asdefined in the first, second, third, fourth, fifth, sixth, eighth,fifteenth, sixteenth, seventeenth, eighteenth, and/or nineteenthembodiments.

In a twenty third embodiment, the invention provides a compoundrepresented by Structural Formula XIII(a) or XIII(b):

or a pharmaceutically acceptable salt thereof; and the variables are asdefined in the first, second, third, fourth, fifth, sixth, eighth,fifteenth, sixteenth, seventeenth, eighteenth, and/or nineteenthembodiments.

In a twenty fourth embodiment, the invention provides a compoundrepresented by Structural Formula XIV:

or a pharmaceutically acceptable salt thereof; and the variables are asdefined in the first, second, third, fourth, fifth, sixth, tenth,fifteenth, sixteenth, seventeenth, eighteenth, and/or nineteenthembodiments.

In a twenty fifth embodiment, the invention provides a compoundaccording to Structural Formula I, II, III, IV, V, VI, VII, VIII, IX, X,XI, XII, XIII(a), XIII(b), XIV, or a pharmaceutically acceptable saltthereof, wherein R³ is isopropyl, tert-butyl, cyclobutyl, cyclopentyl,benzyl, oxetanyl, tetrahydro-2H-pyranyl, or

and the variables are as defined in the first, second, third, fourth,fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth,thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth,nineteenth, twentieth, twenty first, twenty second, twenty third and/ortwenty fourth embodiments. In an alternative embodiment, R³ is isopropylor oxetanyl. In another alternative embodiment, R³ is isopropyl.

In a twenty sixth embodiment, the invention provides a compoundaccording to Structural Formula I, II, III, IV, V, VI, VII, VIII, IX, X,XI, XII, XIII(a), XIII(b), XIV, or a pharmaceutically acceptable saltthereof, wherein R¹ is tert-butyl; and the variables are as defined inthe first, second, third, fourth, fifth, sixth, seventh, eighth, ninth,tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth,seventeenth, eighteenth, nineteenth, twentieth, twenty first, twentysecond, twenty third, twenty fourth, and/or twenty fifth embodiments.

In a twenty seventh embodiment, the invention provides a compoundaccording to Structural Formula III, IV, V, VI, VII, VIII, IX, X, XI,XII, XIII(a), XIII(b), XIV, or a pharmaceutically acceptable saltthereof, wherein R⁴ is

and the variables are as defined in the first, second, third, fourth,fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth,thirteenth, fourteenth, fifteenth, sixteenth, seventeenth, eighteenth,nineteenth, twentieth, twenty first, twenty second, twenty third, twentyfourth, twenty fifth, and/or twenty sixth embodiments. In an alternativeembodiment, R⁴ is

In another alternative embodiment, R⁴ is

In another alternative embodiment, R⁴ is

Still in another alternative embodiment, R⁴ is

Still in another alternative embodiment, R⁴ is

The present invention provides a compound represented by StructuralFormula I′.

In a first embodiment, the invention provides a compound represented byStructural Formula I′:

or a pharmaceutically acceptable salt thereof, wherein:

the thiazole ring is optionally substituted with —F or —Cl;

X⁴ is NR^(a) or O;

X⁵ and X⁶ are each independently NR^(b) or O;

R¹ is (C₁-C₅)alkyl;

R³ is (C₁-C₅)alkyl, —(C₃-C₇)cycloalkyl, or —(CH₂)_(q)heterocyclyl(wherein the heterocycyl is a monocyclic 3-7-membered heterocyclic ringoptionally substituted with one or more occurrences of methyl), orbenzyl (wherein the benzyl ring is optionally substituted with one ormore occurrences of halogen, methoxy, halomethoxy, methyl, halomethyl,or cyano);

each of R^(a), R^(b), and R^(c) is independently hydrogen or methyl;

R^(d) is independently halogen, methoxy, halomethoxy, methyl,halomethyl, or cyano;

m is 0, 1, 2, or 3;

n is 0, 1, or 2; and

q is 0 or 1.

In a second embodiment, the invention provides a compound represented byStructural Formula I′-1:

or a pharmaceutically acceptable salt thereof, and the variables are asdefined in the first embodiment.

In a third embodiment, the invention provides a compound represented byStructural Formula I′-2:

or a pharmaceutically acceptable salt thereof, and the variables are asdefined in the first embodiment.

In a forth embodiment, the invention provides a compound represented byStructural Formula I′-3:

or a pharmaceutically acceptable salt thereof, and the variables are asdefined in the first embodiment.

In a fifth embodiment, the invention provides a compound represented byStructural Formula I′-4:

or a pharmaceutically acceptable salt thereof, and the variables are asdefined in the first embodiment.

In a sixth embodiment, the invention provides a compound according toStructural Formula I′, I′-1, I′-2, I′-3, or I′-4, or a pharmaceuticallyacceptable salt thereof, wherein X⁴ is NH, and the remaining variablesare as defined in the first embodiment.

In a seventh embodiment, the invention provides a compound according toStructural Formula I′, I′-1, I′-2, I′-3, or I′-4, or a pharmaceuticallyacceptable salt thereof, wherein R³ is (C₁-C₄)alkyl, —(C₄-C₆)cycloalkyl,—(CH₂)_(q)heterocyclyl (wherein the heterocycyl is a monocyclic4-6-membered heterocyclic ring optionally substituted with one methyl),or benzyl, and the remaining variables are as defined in the firstand/or sixth embodiments. In one specific embodiment, R³ is isopropyl,tert-butyl, cyclobutyl, cyclopentyl, oxetanyl, benzyl,tetrahydro-2H-pyranyl, or

In another specific embodiment, R³ is isopropyl or oxetanyl.

In an eighth embodiment, the invention provides a compound according toStructural Formula I′, I′-1, I′-2, I′-3, or I′-4, or a pharmaceuticallyacceptable salt thereof, wherein R^(d) is halogen, and m is 0 or 1, andthe remaining variables are as defined in the first, sixth, and/orseventh embodiments. In one specific embodiment,

In a ninth embodiment, the invention provides a compound according toStructural Formula I′, I′-1, I′-2, I′-3, or I′-4, or a pharmaceuticallyacceptable salt thereof, wherein R¹ is tert-butyl, and the remainingvariables are as defined in the first, sixth, seventh, and/or eighthembodiments.

In a tenth embodiment, the invention provides a compound, or apharmaceutically acceptable salt thereof, wherein the compound isselected from the group consisting of:

In an eleventh embodiment, the invention provides a compound representedby Structural Formula II′:

or a pharmaceutically acceptable salt thereof, wherein:

the thiazole ring is optionally substituted with —F or —Cl;

X⁴ is NR^(a) or O;

X⁵ and X⁶ are each independently NR^(b) or O;

R¹ is (C₁-C₅)alkyl;

R⁴ is (C₁-C₄)alkyl, —(C₃-C₇)cycloalkyl, —(CH(R^(c)))_(q)-heterocycyl(wherein the heterocycyl is a monocyclic 3-7-membered heterocyclic ringoptionally substituted with one or more occurrences of methyl),—(CH(R^(c)))_(q)-phenyl (wherein the phenyl ring is optionallysubstituted with one or more occurrences of halogen, methoxy,halomethoxy, methyl, halomethyl, or cyano), or—(CH(R^(c)))_(q)-2-pyridinyl (wherein the 2-pyridinyl ring is optionallysubstituted with one or more occurrences of halogen, methoxy,halomethoxy, methyl, halomethyl, or cyano);

each of R^(a), R^(b), and R^(c) is independently hydrogen or methyl;

n is 0, 1, or 2; and

q is 0 or 1.

In a twelfth embodiment, the invention provides a compound representedby Structural Formula II′-1:

or a pharmaceutically acceptable salt thereof, and the variables are asdefined in the eleventh embodiment.

In a thirteenth embodiment, the invention provides a compoundrepresented by Structural Formula II′-2:

or a pharmaceutically acceptable salt thereof, and the variables are asdefined in the eleventh embodiment.

In a fourteenth embodiment, the invention provides a compound accordingto Structural Formula II′, II′-1 or II′-2, or a pharmaceuticallyacceptable salt thereof, wherein R⁴ is isopropyl, oxetanyl, cyclobutyl,—CH₂-2-pyrrolidinyl, —CH₂—N-methyl-2-pyrrolidinyl, —CH₂-3-piperidinyl,—CH₂-2-pyrazinyl, —CH₂-2-pyrimidinyl, —CH(R^(c))-phenyl, or—CH(R^(c))-2-pyridinyl, and that the phenyl and 2-pyridinyl rings areeach independently and optionally substituted with one or moreoccurrences of halogen, and the remaining variables are as defined inthe eleventh embodiment. In one specific embodiment, R⁴ is

In another specific embodiment, R⁴ is

In a fifteenth embodiment, the invention provides a compound accordingto Structural Formula II′, II′-1 or II′-2, or a pharmaceuticallyacceptable salt thereof, wherein X⁴ is NH, and the remaining variablesare as defined in the eleventh and/or fourteenth embodiments.

In a sixteenth embodiment, the invention provides a compound accordingto Structural Formula II′, II′-1 or II′-2, or a pharmaceuticallyacceptable salt thereof, wherein R¹ is tert-butyl, and the remainingvariables are as defined in the eleventh, fourteenth, and fifteenthembodiments.

In a seventeenth embodiment, the invention provides a compound, or apharmaceutically acceptable salt thereof, wherein the compound isselected from the group consisting of:

Also included are the compounds disclosed in the Exemplification, bothin the pharmaceutically acceptable salt form and in the neutral form.

The term “pharmaceutically acceptable salt” refers to a pharmaceuticalsalt that is, within the scope of sound medical judgment, suitable foruse in contact with the tissues of humans and lower animals withoutundue toxicity, irritation, and allergic response, and is commensuratewith a reasonable benefit/risk ratio. Pharmaceutically-acceptable saltsare well known in the art. For example, S. M. Berge et al. describespharmacologically acceptable salts in J. Pharm. Sci., 1977, 66, 1-19.

Included in the present teachings are pharmaceutically acceptable saltsof the compounds disclosed herein. Compounds having basic groups canform pharmaceutically acceptable salts with pharmaceutically acceptableacid(s). Suitable pharmaceutically acceptable acid addition salts of thecompounds described herein include salts of inorganic acids (such ashydrochloric acid, hydrobromic, phosphoric, metaphosphoric, nitric, andsulfuric acids) and of organic acids (such as acetic acid,benzenesulfonic, benzoic, ethanesulfonic, methanesulfonic, succinic, andtrifluoroacetic acid acids). Compounds of the present teachings withacidic groups such as carboxylic acids can form pharmaceuticallyacceptable salts with pharmaceutically acceptable base(s). Suitablepharmaceutically acceptable basic salts include ammonium salts, alkalimetal salts (such as sodium and potassium salts) and alkaline earthmetal salts (such as magnesium and calcium salts).

Definitions

The term “halo” as used herein means halogen and includes fluoro,chloro, bromo and iodo.

The term “alkyl” used alone or as part of a larger moiety, such as“alkoxy” or “haloalkyl” and the like, means saturated aliphaticstraight-chain or branched monovalent hydrocarbon radical. Unlessotherwise specified, an alkyl group typically has 1-5 carbon atoms, i.e.(C₁-C₅)alkyl. As used herein, a “(C₁-C₅)alkyl” group means a radicalhaving from 1 to 5 carbon atoms in a linear or branched arrangement.Examples include methyl, ethyl, n-propyl, iso-propyl, and the like.

The term “alkoxy” means an alkyl radical attached through an oxygenlinking atom, represented by —O-alkyl. For example, “(C₁-C₄)alkoxy”includes methoxy, ethoxy, propoxy, and butoxy.

The terms “haloalkyl” and “haloalkoxy” means alkyl or alkoxy, as thecase may be, substituted with one or more halogen atoms.

An “alkylene group” is a saturated aliphatic branched or straight-chaindivalent hydrocarbon radical. Unless otherwise specified, an alkylenegroup typically has 2-6 carbon atoms, e.g. (C₂-C₆)alkylene.

The term “alkenyl” means branched or straight-chain monovalenthydrocarbon radical containing at least one double bond. Alkenyl may bemono or polyunsaturated, and may exist in the E or Z configuration.Unless otherwise specified, an alkenyl group typically has 2-6 carbonatoms, i.e., (C₂-C₆)alkenyl. For example, “(C₂-C₄)alkenyl” means aradical having from 2-4 carbon atoms in a linear or branchedarrangement.

The term “cycloalkyl” means a monocyclic saturated hydrocarbon ringsystem. For example, a C₃-C₆ cycloalkyl includes cyclopropyl,cyclobutyl, cyclopentyl and cyclohexyl. Unless otherwise described, a“cycloalkyl” has from three to seven ring carbon atoms.

A bridged cycloalkyl means a bicyclic non-aromatic hydrocarbon ringsystem in which the two rings share at least three adjacent ring carbonatoms. A bridged cycloalkyl typically has 6-12 ring carbon atoms.Examples include, but are not limited to, bicyclo[2.1.1]hexyl,bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, bicyclo[3.2.1]octyl,bicyclo[4.3.1]decyl, bicyclo[3.3.1]nonyl, bornyl, bornenyl, norbornyl,norbornenyl, 6,6-dimethylbicyclo [3.1.1]heptyl, tricyclobutyl, andadamantyl.

The terms “heterocyclyl”, “heterocyclic ring”, and “heterocyclic group”,are used interchangeably herein, and means a saturated or unsaturatednon-aromatic 4-10 membered ring radical containing from 1 to 4 ringheteroatoms, which may be the same or different, selected from N, O, orS. It can be monocyclic, bicyclic or tricyclic (e.g., a fused or bridgedbicyclic or tricyclic ring). Examples of include, but are not limitedto, azetidinyl, morpholinyl, thiomorpholinyl, pyrrolidinonyl,pyrrolidinyl, piperidinyl, piperazinyl, hydantoinyl, valerolactamyl,oxiranyl, oxetanyl, dihydroimidazole, dihydrofuranyl, dihydropyranyl,dihydropyridinyl, dihydropyrimidinyl, dihydrothienyl, dihydrothiophenyl,dihydrothiopyranyl, tetrahydroimidazole, tetrahydrofuranyl,tetrahydropyranyl, tetrahydrothienyl, tetrahydropyridinyl,tetrahydropyrimidinyl, tetrahydrothiophenyl, and tetrahydrothiopyranyl.A heterocyclic ring optionally contains one or more double bonds and/oris optionally fused with one or more aromatic rings (for example,tetrahydronaphthyridine, indolinone, dihydropyrrolotriazole,imidazopyrimidine, quinolinone, dioxaspirodecane).

Examples of 3-7 membered monocyclic heterocyclic ring include, but arenot limited to, azetidinyl, morpholinyl, thiomorpholinyl,pyrrolidinonyl, pyrrolidinyl, piperidinyl, piperazinyl, hydantoinyl,valerolactamyl, oxiranyl, oxetanyl, dihydroimidazole, dihydrofuranyl,dihydropyranyl, dihydropyridinyl, dihydropyrimidinyl, dihydrothienyl,dihydrothiophenyl, dihydrothiopyranyl, tetrahydroimidazole,tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl,tetrahydropyridinyl, tetrahydropyrimidinyl, tetrahydrothiophenyl, andtetrahydrothiopyranyl.

A bridged heterocyclyl means a bicyclic non-aromatic ring systemcontaining from 1 to 4 ring heteroatoms in which the two rings share atleast three adjacent ring atoms. A bridged heterocyclyl typically has6-12 ring atoms. Examples include, but are not limited to, azanorbornyl,quinuclidinyl, isoquinuclidinyl, tropanyl, azabicyclo[3.2.1]octanyl,azabicyclo[2.2.1]heptanyl, 2-azabicyclo[3.2.1]octanyl,azabicyclo[3.2.1]octanyl, azabicyclo[3.2.2]nonanyl,azabicyclo[3.3.0]nonanyl, and azabicyclo [3.3.1]nonanyl.

The terms “heteroaryl”, “heteroaromatic”, “heteroaryl ring”, “heteroarylgroup”, “heteroaromatic ring”, and “heteroaromatic group”, are usedinterchangeably herein. “Heteroaryl” when used alone or as part of alarger moiety as in “heteroaralkyl” or “heteroarylalkoxy”, refers toaromatic ring groups having five to ten ring atoms selected from carbonand at least one (typically 1 to 4, more typically 1 or 2) heteroatoms(e.g., oxygen, nitrogen or sulfur). “Heteroaryl” includes monocyclicrings and polycyclic rings in which a monocyclic heteroaromatic ring isfused to one or more other aromatic or heteroaromatic rings.“Heteroaryl” includes monocyclic and bicyclic ring systems.

“Monocyclic 5-6 membered heteroaromatic ring (or heteroaryl)” means amonocyclic heteroaromatic ring having five or six ring atoms selectedfrom carbon and at least one (typically 1 to 3, more typically 1 or 2)heteroatoms (e.g., oxygen, nitrogen or sulfur). Examples of monocyclic5-6 membered heteroaromatic ring groups include furanyl (e.g.,2-furanyl, 3-furanyl), imidazolyl (e.g., N-imidazolyl, 2-imidazolyl,4-imidazolyl, 5-imidazolyl), isoxazolyl (e.g., 3-isoxazolyl,4-isoxazolyl, 5-isoxazolyl), oxadiazolyl (e.g., 2-oxadiazolyl,5-oxadiazolyl), oxazolyl (e.g., 2-oxazolyl, 4-oxazolyl, 5-oxazolyl),pyrazolyl (e.g., 3-pyrazolyl, 4-pyrazolyl), pyrrolyl (e.g., 1-pyrrolyl,2-pyrrolyl, 3-pyrrolyl), pyridyl (e.g., 2-pyridyl, 3-pyridyl,4-pyridyl), pyrimidinyl (e.g., 2-pyrimidinyl, 4-pyrimidinyl,5-pyrimidinyl), pyridazinyl (e.g., 3-pyridazinyl), thiazolyl (e.g.,2-thiazolyl, 4-thiazolyl, 5-thiazolyl), isothiazolyl, triazolyl (e.g.,2-triazolyl, 5-triazolyl), tetrazolyl (e.g., tetrazolyl), and thienyl(e.g., 2-thienyl, 3-thienyl).

If a group is described as being “substituted,” a non-hydrogensubstituent replaces a hydrogen on a carbon or nitrogen of thesubstituent. Thus, for example, a substituted alkyl is an alkyl whereinat least one non-hydrogen substituent is in the place of a hydrogensubstituent on the alkyl substituent. To illustrate, monofluoroalkyl isalkyl substituted with a fluoro substituent, and difluoroalkyl is alkylsubstituted with two fluoro substituents. It should be recognized thatif there is more than one substitution on a substituent, eachnon-hydrogen substituent can be identical or different (unless otherwisestated). As used herein, many moieties (e.g., alkyl, cycloalkyl, or aheterocyclic ring) are referred to as being either “substituted” or“optionally substituted”. When a moiety is modified by one of theseterms, unless otherwise noted, it denotes that any portion of the moietythat is known to one skilled in the art as being available forsubstitution can be substituted, which includes one or moresubstituents. If more than one substituent is present, then eachsubstituent is independently selected. Such means for substitution arewell-known in the art and/or taught by the instant disclosure. Theoptional substituents can be any substituents that are suitable toattach to the moiety. A person of ordinary skill in the art willrecognize that the compounds and definitions provided do not includeimpermissible substituent patterns (e.g., methyl substituted with 5different groups, and the like). Such impermissible substitutionpatterns are clearly recognized by a person of ordinary skill in theart. When a group is described as being optionally substituted by “oneor more” substituents, it denotes that the group is optionallysubstituted by one, two, three, four, five or six substituents. In oneembodiment, a group is optionally substituted by 1-3 substituents. Inone embodiment, a group is optionally substituted by 1-2 substituents.In one embodiment, a group is optionally substituted by one substituent.

Suitable substituents are those which do not have a significant adverseeffect on the ability of the compound to inhibit RAD51. Where suitablesubstituents are not specifically enumerated, exemplary substituentsinclude, but are not limited to, halo, —CN, alkyl, alkoxy, halomethyl,halomethoxy, (C₁-C₅)alkyl, halo(C₁-C₅)alkyl, (C₁-C₅)alkoxy, —NO₂,—OR^(c′)—NR^(a′)R^(b′), —S(O)_(i)R^(a′), —NR^(a)S(O)_(i)R^(b′),—S(O)_(i)NR^(a′)R^(b′), —C(═O)OR^(a′), —OC(═O)OR^(a′), —C(═S)OR^(a′),O(C═S)R^(a′),

—C(═O)NR^(a′)R^(b′), —NR^(a′)C(═O)R^(b′), —C(═S)NR^(a′)R^(b′),—NR^(a′)C(═S)R^(b′), —NR^(a′)(C═O)OR^(b′), —O(C═O)NR^(a′)R^(b′), —NR^(a)(C═S)OR^(b′), —O(C═S)NR^(a′)R^(b′), —NR^(a′)(C═O)NR^(a′)R^(b′), —NR^(a)(C═S)NR^(a′)R^(b′),

—C(═S)R^(a), —C(═O)R^(a), (C₃-C₆)cycloalkyl, monocyclic heteroaryl andphenyl, wherein the (C₃-C₆)cycloalkyl, monocyclic heteroaryl and phenylsubstituents are optionally and independently substituted with —CH₃,halomethyl, halo, methoxy or halomethoxy. Each R^(a′) and each R^(b′)are independently selected from —H and (C₁-C₅)alkyl, wherein the(C₁-C₅)alkyl group represented by R^(a′)or R^(b′) is optionallysubstituted with hydroxyl or (C₁-C₃)alkoxy; R^(c′) is —H,halo(C₁-C₅)alkyl or (C₁-C₅)alkyl, wherein the (C₁-C₅)alkyl grouprepresented by R^(c) is optionally substituted with hydroxyl or(C₁-C₃)alkoxy; and i is 0, 1, or 2. ═O is also a suitable substituentfor alkyl, cycloalkyl, and a heterocyclic ring.

Compounds having one or more chiral centers can exist in variousstereoisomeric forms. Stereoisomers are compounds that differ only intheir spatial arrangement. Stereoisomers include all diastereomeric,enantiomeric, and epimeric forms as well as racemates and mixturesthereof.

The term “geometric isomer” refers to cyclic compounds having at leasttwo substituents, wherein the two substituents are both on the same sideof the ring (cis) or wherein the substituents are each on opposite sidesof the ring (trans). When a disclosed compound is named or depicted bystructure without indicating stereochemistry, it is understood that thename or the structure encompasses one or more of the possiblestereoisomers, or geometric isomers, or a mixture of the encompassedstereoisomers or geometric isomers.

When a geometric isomer is depicted by name or structure, it is to beunderstood that the named or depicted isomer exists to a greater degreethan another isomer, that is that the geometric isomeric purity of thenamed or depicted geometric isomer is greater than 50%, such as at least60%, 70%, 80%, 90%, 99%, or 99.9% pure by weight. Geometric isomericpurity is determined by dividing the weight of the named or depictedgeometric isomer in the mixture by the total weight of all of thegeometric isomers in the mixture.

Racemic mixture means 50% of one enantiomer and 50% of is correspondingenantiomer. When a compound with one chiral center is named or depictedwithout indicating the stereochemistry of the chiral center, it isunderstood that the name or structure encompasses both possibleenantiomeric forms (e.g., both enantiomerically-pure,enantiomerically-enriched or racemic) of the compound. When a compoundwith two or more chiral centers is named or depicted without indicatingthe stereochemistry of the chiral centers, it is understood that thename or structure encompasses all possible diastereomeric forms (e.g.,diastereomerically pure, diastereomerically enriched and equimolarmixtures of one or more diastereomers (e.g., racemic mixtures) of thecompound.

Enantiomeric and diastereomeric mixtures can be resolved into theircomponent enantiomers or stereoisomers by well-known methods, such aschiral-phase gas chromatography, chiral-phase high performance liquidchromatography, crystallizing the compound as a chiral salt complex, orcrystallizing the compound in a chiral solvent. Enantiomers anddiastereomers also can be obtained from diastereomerically- orenantiomerically-pure intermediates, reagents, and catalysts bywell-known asymmetric synthetic methods.

When a compound is designated by a name or structure that indicates asingle enantiomer, unless indicated otherwise, the compound is at least60%, 70%, 80%, 90%, 99% or 99.9% optically pure (also referred to as“enantiomerically pure”). Optical purity is the weight in the mixture ofthe named or depicted enantiomer divided by the total weight in themixture of both enantiomers.

When the stereochemistry of a disclosed compound is named or depicted bystructure, and the named or depicted structure encompasses more than onestereoisomer (e.g., as in a diastereomeric pair), it is to be understoodthat one of the encompassed stereoisomers or any mixture of theencompassed stereoisomers is included. It is to be further understoodthat the stereoisomeric purity of the named or depicted stereoisomers atleast 60%, 70%, 80%, 90%, 99% or 99.9% by weight. The stereoisomericpurity in this case is determined by dividing the total weight in themixture of the stereoisomers encompassed by the name or structure by thetotal weight in the mixture of all of the stereoisomers.

Pharmaceutical Compositions

The compounds disclosed therein are RAD51 inhibitors. The pharmaceuticalcomposition of the present invention comprises one or more RAD51inhibitors, or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier or diluent.

“Pharmaceutically acceptable carrier” and “pharmaceutically acceptablediluent” refer to a substance that aids the formulation and/oradministration of an active agent to and/or absorption by a subject andcan be included in the compositions of the present disclosure withoutcausing a significant adverse toxicological effect on the subject.Non-limiting examples of pharmaceutically acceptable carriers and/ordiluents include water, NaCl, normal saline solutions, lactatedRinger's, normal sucrose, normal glucose, binders, fillers,disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions(such as Ringer's solution), alcohols, oils, gelatins, carbohydratessuch as lactose, amylose or starch, fatty acid esters,hydroxymethycellulose, polyvinyl pyrrolidine, and colors, and the like.Such preparations can be sterilized and, if desired, mixed withauxiliary agents such as lubricants, preservatives, stabilizers, wettingagents, emulsifiers, salts for influencing osmotic pressure, buffers,coloring, and/or aromatic substances and the like that do notdeleteriously react with or interfere with the activity of the compoundsprovided herein. One of ordinary skill in the art will recognize thatother pharmaceutical excipients are suitable for use with disclosedcompounds.

The pharmaceutical compositions of the present teachings optionallyinclude one or more pharmaceutically acceptable carriers and/or diluentstherefor, such as lactose, starch, cellulose and dextrose. Otherexcipients, such as flavoring agents; sweeteners; and preservatives,such as methyl, ethyl, propyl and butyl parabens, can also be included.More complete listings of suitable excipients can be found in theHandbook of Pharmaceutical Excipients (5^(th) Ed., Pharmaceutical Press(2005)). A person skilled in the art would know how to prepareformulations suitable for various types of administration routes.Conventional procedures and ingredients for the selection andpreparation of suitable formulations are described, for example, inRemington's Pharmaceutical Sciences (2003-20th edition) and in TheUnited States Pharmacopeia: The National Formulary (USP 24 NF19)published in 1999. The carriers, diluents and/or excipients are“acceptable” in the sense of being compatible with the other ingredientsof the pharmaceutical composition and not deleterious to the recipientthereof.

Methods of Treatment

The present invention provides a method of treating a subject with adisease which can be ameliorated by inhibition of RAD51, byadministering to the subject an effective amount of one or moredisclosed compounds, or a pharmaceutically acceptable salt thereof, orthe corresponding pharmaceutical composition. Diseases which can beameliorated by inhibition of RAD51 include treating cancer, autoimmunedisease, immune deficiency, or neurodegenerative disease.

In one aspect, described herein is a method of treating cancer,autoimmune disease, immune deficiency, or neurodegenerative disease, themethod comprising administering a therapeutically effective dose of acomposition as described herein, e.g., a composition comprising acompound of the present invention, to a subject in need of treatment forcancer, autoimmune disease, immune deficiency, or neurodegenerativedisease.

In some embodiments, the subject can be a subject determined to have anincreased level of DNA damage occurring in one or more cell typesrelative to a reference level. As used herein, “DNA damage” refers tobreaks, nicks, and mutations of the DNA present in a cell. In someembodiments, the DNA damage can comprise one or more of single-strandbreaks (e.g., “nicks”), double strand breaks (DSBs), and mutations. Insome embodiments, the DNA damage can be one or more DSBs. As usedherein, “mutation” refers to a change or difference in the geneticmaterial of a cell as compared to a reference wildtype cell, e.g. adeletion, an insertion, a SNP, a gene rearrangement, and/or theintroduction of an exogenous gene or sequence.

In some embodiments, the subject can be determined to have an increasedlevel of DNA damage if the subject is determined to have an increasedlevel and/or activity of a DNA damage process or DNA editing enzyme. Asused herein, “DNA damage process” refers to any activity or process in acell which causes one or more types of DNA damage to occur.

In some embodiments, an increased level of DNA damage can be anincreased level of mutations, e.g., by determining the overall mutationstatus in all or a portion of the genome of a cell. An overall mutationstatus at least 2% greater, e.g. 2% greater or more, 3% greater or more,5% greater or more, 10% greater or more, or 20% greater or more than theoverall mutation status in a reference cell can be indicative of anincreased, elevated, and/or significant level of a DNA editing enzymeactivity. In some embodiments, the level of hyper mutations can bedetermined. In some embodiments, the overall mutation status in thewhole genome or a portion thereof can be determined using FISH, wholegenome sequencing, high throughput sequencing, exome sequencing,hybridization, and/or PCR. In some embodiments the activity of a DNAediting enzyme can be measured by determining the level ofhypermutations in the specific target genes including, but not limitedto IGH, BCL6, MYC, BCL1 1A, CD93, PIM1 and/or PAX5. In certainembodiments the DNA editing enzyme is AID. In some embodiments, a levelof mutation in specific target genes including IGH, BCL6, MYC, BCL1 1A,CD93, PIM1 and/or PAX5 which is at least 2% greater, e.g. 2% greater ormore, 3% greater or more, 5% greater or more, 10% greater or more, or20% greater or more than the level of mutation in IGH, BCL6, MYC, BCL11A, CD93, PIM1 and/or PAX5 in a reference cell can be indicative of anincreased, elevated, and/or significant level of AID activity.

In some embodiments, an increased level of DNA damage can be anincreased level of double strand breaks (DSBs). The level of DSBs can bedetermined, by way of non-limiting example, by karyotyping, by γ-H2AXfoci formation, and/or by using FISH analysis to detect DNA doublestrand breaks, e.g. DNA breakage detection fish (DBD-FISH) (Volpi andBridger, BioTechniques, Vol. 45, No. 4, October 2008, pp. 385-409).

In some embodiments, an increased level of DNA damage can be anincreased level of single strand breaks. The level of single-strandbreaks in DNA can be determined, by way of non-limiting example, byCOMET assays, FISH, or the use of single-strand break-specific probes.Detection of DNA breaks, both single and double-stranded is known in theart and described further, at, e.g., Kumari et al. EXCLI Journal 20097:44-62 and Motalleb et al. Research Journal of Applied Sciences,Engineering and Technology. 2012 4: 1888-1894; each of which isincorporated by reference herein in its entirety.

In some embodiments, an increased level of activity of a DNA damageprocess can comprise an increased level and/or activity of a DNA editingenzyme. In some embodiments, the technology described herein is directedto treating cells having an active DNA editing enzyme with a compound ofthe present invention. In some embodiments, the technology describedherein is directed to treating cells having an increased level and/oractivity of a DNA editing enzyme with a compound of the presentinvention. As used herein, “DNA editing enzyme” refers to an enzymewhich normally catalyzes the mutation, exchange or excision of DNAsegments, particularly enzymes which can generate or promote thegeneration of point mutations, DNA single strand breaks, DNAdouble-strand breaks or protein-DNA adducts. A DNA editing enzyme, asreferred to herein, is not necessarily site-specific in its action.Similarly, it is not necessarily cell specific. In some embodiments, thecell is a B cell expressing a detectable amount of such an enzyme.

Non-limiting examples of DNA editing enzymes include, but are notlimited to Recombination Activating Gene 1 (RAG1; NCBI Gene ID: 5896),Recombination Activating Gene 1 (RAG2; NCBI Gene ID: 5897),Sporulation-specific protein 11 (SPO1 1; NCBI Gene ID: 23626), APOBECfamily members a Type 1 Topoisomerase; a Type 2 Topoisomerase; and/orAID. In some embodiments, the DNA editing enzyme can be AID.

In some embodiments, the DNA editing enzyme can be a member of theAPOBEC (apolipoprotein B mRNA editing enzyme, catalyticpolypeptide-like) family. As used herein “APOBEC family” refers to afamily of cytidine deaminase enzymes having an N-terminal zinc-dependentcytidine deaminase catalytic domain comprising and a C-terminalpseudocatalytic domain. Non-limiting examples of APOBEC family membersinclude AID, APOBEC 1 (e.g., NCBI Gene ID: 339), APOBEC2 (e.g., NCBIGene ID: 10930), APOBEC3A (e.g., NCBI Gene ID: 200315), APOBEC3C (e.g.,NCBI Gene ID: 27350), APOBEC3E (e.g., NCBI Gene ID: 140564), APOBEC3F(e.g., NCBI Gene ID:200316), APOBEC3G (e.g., NCBI Gene ID: 60489),APOBEC3H (e.g., NCBI Gene ID: 164668), and APOBEC4 (e.g., NCBI Gene ID:403314).

In some embodiments, the DNA editing enzyme can be a Type 1topoisomerase. In some embodiments, the DNA editing enzyme can be a Type2 topoisomerase. Topoisomerases generate breaks in DNA to help uncoil orrelax the strand. Type II topoisomerases hydrolyze ATP to generate DSBcuts, while Type I topoisomerases generate single-stranded breaks.Non-limiting examples of Type II topoisomerases can includetopoisomerase II (e.g., NCBI Gene ID: 7153 and 7155). Non-limitingexamples of Type I topoisomerases can include topoisomerase I (e.g.,NCBI Gene ID: 7150).

Embodiments of the technology described herein are based on thediscovery that the compounds described herein can inhibit DNA repairmechanisms, e.g., homologous repair. Activation-induced cytidinedeaminase (AID, or AICDA, also known as ARP2, CDA2 or HIGM2), aDNA-editing enzyme that is a member of the apolipoprotein B mRNA editingenzymes, catalytic polypeptide-like (APOBEC), will cause widespreadgenomic breaks and cell death in cells with diminished homologousrecombination ability (e.g. cells with diminished DNA double strandbreak repair abilities). Accordingly, provided herein is a method ofcausing cell death comprising detecting increased expression of aDNA-editing enzyme (e.g. AID) in a cell and thereafter contacting thecell with a compound of the present invention; thereby resulting in celldeath. Accordingly, provided herein is a method of causing cell deathcomprising increasing expression of a DNA-editing enzyme (e.g. AID) in acell and thereafter contacting the cell with a compound of the presentinvention; thereby resulting in cell death. Accordingly, provided hereinis a method of causing cell death comprising administering to a cell atherapeutically effective amount of a DNA editing enzyme (e.g. AID) andthereafter contacting the cell with a compound of the present invention;thereby resulting in cell death.

AID, encoded by the AICDA gene (NCBI Gene ID: 57379), is required forproper B-cell function and is most prominently expressed in centroblastB-cells. The protein is involved in somatic hypermutation, geneconversion, and class-switch recombination of immunoglobulin genes. AIDis normally expressed almost exclusively in antigen-activated germinalcenter B-cells, where it initiates immunoglobulin isotype classswitching (Manis et al. 2002, Trends Immunol, 23, 31-39; Chaudhuri andAlt, Nat Rev Immunol, 2004, 4, 541-552; Longerich et al., Curr OpinImmunol, 2006, 18, 164-174; Chaudhuri et al., Adv Immunol 2007, 94,157-214). AID is required for somatic hypermutation and immunoglobulinclass switching in activated B cells. AID expression is regulated byCD40 ligand, B-cell receptor, IL4R, or Toll-like receptor stimulation(Crouch et al., J Exp Med 2007 204: 1145-1156; Muramatsu et al., J BiolChem 1999 274: 18470-6). After activation, AID is transientlyupregulated, induces point mutations or DNA double strand breaks in asequence nonspecific manner within immunoglobulin genes, and is thendownregulated (Longerich et al., Curr Opin Immunol, 2006, 18, 164-176;Chaudhuri et al., Adv Immunol 2007, 94, 157-214). Overall, AID is activein only a tiny population of normal cells (antigen-activated B-cells) atany given time. The genomic rearrangements and mutations controlled byAID lead to the development of antigen-recognition diversity, receptorediting and lymphoid effector function required for functional adaptiveimmunity (Mills, et al. Immunol Rev 2003 194:77-95). Recently it hasbeen reported that AID has off-target point mutation activities (Liu, M.et al., Nature 2008, 451, 841-845; Liu and Schatz, Trends Immunol. 2009,30, 173-181; Perez-Duran et al., Carcinogenesis. 2007, 28(12):2427-33).Robbiani et al. has reported off-target activities of AID in B− cells,especially c-myc/IgH translocations (Robbiani et al., Mol Cell 2009,36(4):631-41). AID expression accelerates the rate of tumor developmentin Bcl6 transgenic mice (Pasqualucci et al., 2008, Nat. Genet. 40,108-112). However, deregulated AID does not necessarily cause malignancyor translocation-associated cancer on its own in B cells (Muto et al.,2006, Proc. Natl. Acad. Sci. USA 103, 2752-2757; Okazaki et al., 2003,J. Exp. Med. 197, 1173-1181; Shen et al., 2008, Mol. Immunol. 45,1883-1892). In addition, despite its obligate role in c-myc/IgHtranslocation, AID is not required for the development of plasmacytosisor plasmacytoma in IL-6 transgenic or pristane-treated mice,respectively (Kovalchuk et al., 2007, J. Exp. Med. 204, 2989-3001;Ramiro et al., 2004, J. Exp. Med. 200, 1103-1110). However, most human Bcell lymphoma-associated translocations do not involve c-myc, and manydo not involve Ig genes (Kuppers, 2005, Oncogene 20, 5580-5594).

Overexpression of AID has been reported in chronic lymphocytic leukemia(CLL) (Hancer et al. Leuk Lymphoma. 2011 January; 52(1):79-84; Heintelet al., Leukemia. 2004 April; 18(4):756-62). Further, AID expression hasbeen shown to be correlated with blast crisis B lineage leukemia andtherapy resistance in myeloid leukemia and to be associated withgenerally poor prognosis in chronic B lymphocytic leukemia (Mao et al.,Br J Dermatol 2001, 145: 117-122; Chaudhuri et al., Nature 2004,430:992-8). Further expression of AID in tumor cells from a variety ofcancers has been reported including but not limited to lung, breast,gastric, colon, intestinal, liver cancer and choriangiocarcinoma (Greeveet al., Blood 2003, 1010, 3574-3580; Feldhahn et al., J Exp Med 2007,204, 1157-1166; Kotani et al., PNAS USA 2007, 104, 1616-1620; Engels etal., 2008, Appl Immunohistochem Mol Morphol 16, 521-529; Klemm et al.,2009, Cancer Cell 6, 232-245; Palacios et al., 2010, Blood 115(22),4488-4496; Leuenberger et al., 2009, Mod Pathol 32, 177-186; Gruber etal., 2010, Cancer Res 70, 7411-7420; inflammatory cancer (Marusawa 2008,Int J Biochem Cell Biol. 40, 399-402); follicular lymphoma (Hardianti etal., 2004, Leukemia 18, 826-831; Shikata et al., 2012, Cancer Sci.103(3):415-21); thyroid cancer (Qiu et al. 2012, Mod Pathol 25(1),36-45); breast cancer (Borchert et al. 2011, BMC Cancer 11:347);Marusawa, et al., 2011, Adv Immunol 111: 109-41; Zhang et al. 2012, HumPathol 43(3):423-34; Komori et al., 2008, Hepatology 47(3):888-896;Hockley 2010, Leukemia 24(5): 1084-6; adult T-cell leukemia (Nakamura etal., 2011, Br J Dermatol. 165(2):437-9). All of the references in theforegoing paragraph are incorporated by reference herein in theirentireties.

Elevated levels of AID have been reported in arthritis (Xu et al. Scand.J. Immunol. 2009, 296, 2033-6) and in the MRL/Fas(lpr/lpr) mouse lupusmodel (White et al. 2011, Autoimmunity 44(8), 585-98). All of thereferences in the foregoing paragraph are incorporated by referenceherein in their entireties.

When DSB repair is inhibited, the extent of the DSBs generated by AID ismuch higher than previously suspected and the extent of genomic damageis so severe as to result in cell death. Accordingly, in one embodimentof the technology described herein, there is provided a method oftreatment comprising; (a) selecting a subject having cells that expresselevated levels of activation-induced cytidine deaminase (AID); and (b)administering a therapeutically effective amount of an inhibitor ofdouble strand break repair (e.g. a compound of the present invention) tothe subject; wherein an elevated level of AID is a level of AID that ishigher than the level of AID in cells of the same type from a healthyindividual. In some embodiments, the cells expressing elevated levels ofAID are B cells. In some embodiments, the B cell expressing elevatedlevels of AID is a cancerous B cells or a B cell associated withautoimmune disease. In some embodiments, the subject can be a humansubject.

Methods provided herein treat cancers and/or autoimmune disorders byinhibiting DNA double strand break repair. This inhibition proves lethalto cells expressing AID, as AID generates widespread genomic breaks, andthe treatment with a double strand break repair inhibitor prevents therepair of these lesions which are being generated by the cell itself.This results in cell death in the subject which is specific to the cellsexpressing AID, e.g. cancerous B cells and/or autoimmune cells.Accordingly, as described herein, in one embodiment there is a provideda treatment paradigm that selectively induces self-destruction ofcertain diseased cells, while reducing the unintended side effects inhealthy tissues.

In some embodiments, an increased level and/or activity of a DNA editingenzyme can be an increased level of DNA editing enzyme mRNA. mRNA levelscan be assessed using, e.g., biochemical and molecular biologytechniques such as Northern blotting or other hybridization assays,nuclease protection assay, reverse transcription (quantitative RT-PCR)techniques, RNA-Seq, high throughput sequencing and the like. Suchassays are well known to those in the art. In one embodiment, nuclear“run-on” (or “run-off) transcription assays are used (see e.g. Methodsin Molecular Biology, Volume: 49, Sep. 27, 1995, Page Range: 229-238).Arrays can also be used; arrays, and methods of analyzing mRNA usingsuch arrays have been described previously, e.g. in EP0834575,EP0834576, WO96/31622, U.S. Pat. No. 5,837,832 or WO98/30883. WO97/10365provides methods for monitoring of expression levels of a multiplicityof genes using high density oligonucleotide arrays.

In some embodiments, a subject can be determined to have an increasedlevel of DNA damage occurring in one or more cell types relative to areference level if the subject has been exposed to an agent that isknown to cause such DNA damage. Non-limiting examples of such agents caninclude a viral infection with a DNA integrating virus (e.g.adeno-associated virus, retrovirus, human T-lymphotropic virus, HIV-1,oncovirus, hepatitis virus, hepatitis B virus); DNA damaging chemicals(e.g. acetaldehyde, polycyclic aromatic hydrocarbons, benzenes,nitrosamines, tobacco smoke, aflatoxin, and the like); DNA damagingchemotherapeutic agents (e.g. bleomycin, mitomycin, nitrogen mustards(e.g. mechlorethamine, cyclophosphamide, melphalan, chlorambucil,ifosfamide and busulfan), nitrosoureas (e.g., N-Nitroso-N-methylurea(MNU), carmustine (BCNU), lomustine (CCNU) and semustine (MeCCNU),fotemustine and streptozotocin), tetrazines (e.g., dacarbazine,mitozolomide and temozolomide), aziridines (e.g., thiotepa, mytomycinand diaziquone (AZQ)), cisplatins (e.g., cisplatin, carboplatin andoxaliplatin) procarbazine and hexamethylmelamine); and ionizing orultraviolet radiation. Exposure to such agents can be the result of anaccident, infection and/or environmental exposure or the result of atherapeutic administration of such agents.

In some embodiments, the increased level of DNA damage can be occurringin a cell type affected by the cancer, autoimmune disease, and/orneurodegenerative disease. In some embodiments, the subject isdetermined to have an increased level of DNA damage occurring in a cellselected from the group consisting of: a cancer cell; an immune systemcell; or a nervous system cell.

In some embodiments, the DNA editing enzyme can be AID. In someembodiments, the level of AID can be the level of AID in a blood cell.In some embodiments, the level of AID can be the level of AID in a Bcell.

In some embodiments, an increased level of AID can be a detectable levelof AID, e.g., as described below herein.

In some embodiments, the subject can be a human subject.

Methods provided herein treat cancers and/or autoimmune disorders byinhibiting DNA double strand break repair. This inhibition proves lethalto cells expressing AID, as AID generates widespread genomic breaks, andthe treatment with a double strand break repair inhibitor prevents therepair of these lesions which are being generated by the cell itself.This results in cell death in the subject which is specific to the cellsexpressing AID, e.g. cancerous B cells and/or autoimmune cells.Accordingly, as described herein, in one embodiment there is a provideda treatment paradigm that selectively induces self-destruction ofcertain diseased cells, while reducing the unintended side effects inhealthy tissues.

Methods of defecting cancers in patients with increased levels of DNAdamage or increased levels of DNA editing enzymes are disclosed inWO2016/094897, incorporated herein by reference.

In certain embodiments, the cancer to be treated is a type with highexpression of a DNA editing enzyme. In certain embodiments, the cancerto be treated is a B-cell neoplasm.

Another embodiment is a method of treating a cancer by administering tothe subject an effective amount of one or more disclosed compounds, or apharmaceutically acceptable salt thereof, or the correspondingpharmaceutical composition. In one aspect, the cancer is selected fromthe group consisting of lymphoma, leukemia, and a plasma cell neoplasm.In another aspect, the cancer selected from the group consisting ofcarcinoma and sarcoma.

In certain embodiments, the cancer to be treated is a lymphoma.Lymphomas which can be treated by the disclosed methods includeNon-Hodgkin's lymphoma; Burkitt's lymphoma; small lymphocytic lymphoma;lymphoplasmacytic lymphoma; MALT lymphoma; follicular lymphoma; diffuselarge B-cell lymphoma; and T-cell lymphoma.

Lymphoma is a malignancy in the lymphatic cells of the immune system(e.g. B cells, T cells, or natural killer (NK) cells). Lymphomas oftenoriginate in the lymph nodes and present as solid tumors. They canmetastasize to other organs such as the brain, bone, or skin. Extranodalsites are often located in the abdomen. Lymphomas are closely related tothe lymphoid leukemia and in some cases a particular form of cancer iscategorized as both a lymphoma and a leukemia.

Leukemias which can be treated by the disclosed methods include acutelymphoblastic leukemia (ALL); Burkitt's leukemia; B-cell leukemia;B-cell acute lymphoblastic leukemia; chronic lymphocytic leukemia (CLL);acute myelogenous leukemia (AML); chronic myelogenous leukemia (CML);and T-cell acute lymphoblastic leukemia (T-ALL).

In certain embodiments the cancer to be treated is B-cell neoplasms,B-cell leukemia, B-cell acute lymphoblastic leukemia, chroniclymphocytic leukemia, chronic myelogenous leukemia, Burkitt's leukemia,acute myelogenous leukemia and/or T-ALL. The maturation of B cells mosttypically ceases or substantially decreases when the foreign antigen hasbeen neutralized. Occasionally, however, proliferation of a particular Bcell will continue unabated; such proliferation can result in a cancerreferred to as “B-cell lymphoma” or a “B-cell leukemia.” In certainembodiments the cancer to be treated is chronic lymphocytic leukemia(CLL) or chronic myelogenous leukemia (CML).

In certain embodiments the cancer to be treated is a plasma cellneoplasm. Examples for plasma cell neoplasms include multiple myeloma;plasma cell myeloma; plasma cell leukemia and plasmacytoma.

Carcinomas which can be treated by the disclosed methods include coloncancer; liver cancer; gastric cancer; intestinal cancer; esophagealcancer; breast cancer; ovarian cancer; head and neck cancer; lungcancer; and thyroid cancer.

Sarcomas which can be treated by the disclosed methods include softtissue sarcoma and bone sarcoma.

Any cancer characterized by high levels of DNA damage and/or DNA editingenzyme expression can be treated with a compound as described herein,e.g. a compound of the present invention. For example, sarcomas,epithelial cell cancer (carcinomas), colon cancer, gastric cancer,intestinal cancer, liver cancer, hepatocellular cancer, breast cancer,thyroid cancer, esophageal cancer, lung cancer, brain cancer, head andneck cancer, melanoma, renal cancer, prostate cancer, hemangioma,rhabdomyosarcoma, chondrosarcoma, osteosarcoma, fibrosarcoma andcholangiocarcinoma may be characterized by high levels of a DNA editingenzyme expression, e.g. AID. In certain embodiments the cancer to betreated is colon cancer, liver cancer, gastric cancer, intestinalcancer, breast cancer, lung cancer, thyroid cancer and/orcholangiocarcinoma.

Specific cancers that can be treated by the disclosed methods includecancer of the bladder, blood, bone, bone marrow, brain, breast, colon,esophagus, gastrointestine, gum, head, kidney, liver, lung, nasopharynx,neck, ovary, prostate, skin, stomach, testis, tongue, or uterus. Inaddition, the cancer may specifically be of the following histologicaltype, though it is not limited to these: neoplasm, malignant; carcinoma;carcinoma, undifferentiated; giant and spindle cell carcinoma; sarcomas;small cell carcinoma; papillary carcinoma; squamous cell carcinoma;lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma;transitional cell carcinoma; papillary transitional cell carcinoma;adenocarcinoma; gastrinoma, malignant; cholangiocarcinoma;hepatocellular carcinoma; combined hepatocellular carcinoma andcholangiocarcinoma; trabecular adenocarcinoma; adenoid cystic carcinoma;adenocarcinoma in adenomatous polyp; adenocarcinoma, familial polyposiscoli; solid carcinoma; carcinoid tumor, malignant; branchiolo-alveolaradenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma;acidophil carcinoma; oxyphilic adenocarcinoma; basophil carcinoma; clearcell adenocarcinoma; granular cell carcinoma; follicular adenocarcinoma;papillary and follicular adenocarcinoma; nonencapsulating sclerosingcarcinoma; adrenal cortical carcinoma; endometroid carcinoma; skinappendage carcinoma; apocrine adenocarcinoma; sebaceous adenocarcinoma;ceruminous adenocarcinoma; mucoepidermoid carcinoma; cystadenocarcinoma;papillary cystadenocarcinoma; papillary serous cystadenocarcinoma;mucinous cystadenocarcinoma; mucinous adenocarcinoma; signet ring cellcarcinoma; infiltrating duct carcinoma; medullary carcinoma; lobularcarcinoma; inflammatory carcinoma; paget's disease, mammary; acinar cellcarcinoma; adenosquamous carcinoma; adenocarcinoma w/squamousmetaplasia; thymoma, malignant; ovarian stromal tumor, malignant;thecoma, malignant; granulosa cell tumor, malignant; androblastoma,malignant; sertoli cell carcinoma; leydig cell tumor, malignant; lipidcell tumor, malignant; paraganglioma, malignant; extra-mammaryparaganglioma, malignant; pheochromocytoma; glomangiosarcoma; malignantmelanoma; amelanotic melanoma; superficial spreading melanoma; maligmelanoma in giant pigmented nevus; epithelioid cell melanoma; bluenevus, malignant; sarcoma; fibrosarcoma; fibrous histiocytoma,malignant; myxosarcoma; liposarcoma; leiomyosarcoma; rhabdomyosarcoma;embryonal rhabdomyosarcoma; alveolar rhabdomyosarcoma; stromal sarcoma;mixed tumor, malignant; mullerian mixed tumor; nephroblastoma;hepatoblastoma; carcinosarcoma; mesenchymoma, malignant; brenner tumor,malignant; phyllodes tumor, malignant; synovial sarcoma; mesothelioma,malignant; dysgerminoma; embryonal carcinoma; teratoma, malignant;struma ovarii, malignant; choriocarcinoma; mesonephroma, malignant;hemangiosarcoma; hemangioendothelioma, malignant; Kaposi's sarcoma;hemangiopericytoma, malignant; lymphangiosarcoma; osteosarcoma;juxtacortical osteosarcoma; chondrosarcoma; chondroblastoma, malignant;mesenchymal chondrosarcoma; giant cell tumor of bone; Ewing's sarcoma;odontogenic tumor, malignant; ameloblastic odontosarcoma; ameloblastoma,malignant; ameloblastic fibrosarcoma; pinealoma, malignant; chordoma;glioma, malignant; ependymoma; astrocytoma; protoplasmic astrocytoma;fibrillary astrocytoma; astroblastoma; glioblastoma; oligodendroglioma;oligodendroblastoma; primitive neuroectodermal; cerebellar sarcoma;ganglioneuroblastoma; neuroblastoma; retinoblastoma; olfactoryneurogenic tumor; meningioma, malignant; neurofibrosarcoma;neurilemmoma, malignant; granular cell tumor, malignant; malignantlymphoma; Hodgkin's disease; hodgkin's; paragranuloma; malignantlymphoma, small lymphocytic; malignant lymphoma, large cell, diffuse;malignant lymphoma, follicular; mycosis fungoides; other specifiednon-Hodgkin's lymphomas; malignant histiocytosis; multiple myeloma; mastcell sarcoma; immunoproliferative small intestinal disease; leukemia;lymphoid leukemia; plasma cell leukemia; erythroleukemia; lymphosarcomacell leukemia; myeloid leukemia; basophilic leukemia; eosinophilicleukemia; monocytic leukemia; mast cell leukemia; megakaryoblasticleukemia; myeloid sarcoma; and hairy cell leukemia.

In another embodiment for the disclosed method, the cancer ischaracterized by mutations in the mutS homologues (e.g., MSH2, MSH3, andMSH6), mutL homologues (e.g. MLH1), or mismatch repair endonucleasePMS2. Mutations are changes in the genetic code. They include pointmutations and frameshift mutations. In a point mutation, one nucleotideis swapped out for another. Therefore, the mutation occurs at a singlepoint or location within the DNA strand. Frameshift mutations are due toeither insertions or deletions of nucleotides. This causes the entireDNA strand to elongate or to shrink in size. Thus, frameshift mutationsmay alter all of the codons that occur after the deletion or insertion.The mutations referred to herein include, but are not limited to,insertions, deletions, duplications, inversions, or other recognizedpoint mutations. It has now been found that RAD51 inhibitors areparticularly effective in treating cancers with mutations in MSH (e.g.MSH6), MLH, or PMS2.

MutS Homolog 2 (MSH2) is a protein that in humans is encoded by the MSH2gene, which is located on chromosome 2. MSH2 is a tumor suppressor geneand more specifically a caretaker gene that codes for a DNA mismatchrepair (MMR) protein, MSH2, which forms a heterodimer with MSH6 to makethe human MutSα mismatch repair complex. It also dimerizes with MSH3 toform the MutSβ DNA repair complex. MSH2 is involved in many differentforms of DNA repair, including transcription-coupled repair, homologousrecombination, and base excision repair. Examples of the mutations inMSH2 include, but are not limited to, g.47630253_47630254del,g.47702411_47702421del, g.47709913_47709915inv, g.47635629_47635634del,g.47637227_47637236dup, g.47639550_47639561del,g.(?_47630206)_(47710367_?)del, g.(?_47630206)_(47643569_47656880)del,g.47630263_47643568del, g.(?_47630206)_(47657081_47672686)del,g.47630263_47657080del, g.(?_47630206)_(47672797_47690169)del,g.47630263_47672796del, g.(?_47630206)_(47672797_47690169)del,g.(?_47630206)_(47693948_47698103)del, g.47630263_47693947del,g.(?_47630206)_(47698202_47702163)del,g.(?_47630206)_(47630542_47635539)del,g.(?_47630206)_(47708011_47709917)del,g.(?_47630206)_(47635695_47637232)del,g.(?_47630206)_(47635695_47637232)del,g.(?_47630206)_(47637512_47639552)del,g.(?_47630206)_(47639700_47641407)del,g.(?_47630206)_(47641558_47643434)del, g.47618487_47650860delins(155),g.47628578_47638433del, g.47595033_47662777del, g.47583175_47667707del,g.47625602_47636880del, g.47554933_47699909del, g.47629508_47649552del,g.47629375_47651274del, g.(?_47630206)_(47630542_47635539)del,g.(?_47630206)_(47635695_47637232)del, g.47643509_47643510del,g.47643529_47643530dup, g.47656746_47657199dup, g.47656661_47663325del,g.(47643569_47656880)_(47710367_?)del,g.(47643569_47656880)_(47710367_?)del, g.47656881_47657080del,g.(47643569_47656880)_(47657081_47672686)del,g.(47643569_47656880)_(47657081_47672686)del,g.(47643569_47656880)_(47657081_47672686)del,g.(47643569_47656880)_(47657081_47672686)dup,g.(47643569_47656880)_(47657081_47672686)dup,g.(47643569_47656880)_(47672797_47690169)del,g.(47643569_47656880)_(47693948_47698103)del, g.47656881_47693947dell,g.(47643569_47656880)_(47702410_47703505)del,g.47656881_47656882ins(173), g.47656901_47656902insA, g.47656903del,g.47656912del, g.47630440del, g.47656923del, g.47656931_47656932dup,g.47656943del, g.47656943_47656949delinsCCCAGA, g.47656948dup,g.47656996dup, g.47657000_47657001dup, g.47630449del, g.47657007dup,g.47657008del, g.47657020_47657023dup, g.47657025_47657026del,g.47657026dup, g.47657030_47657031del, g.47657047_47657050del,g.47657053del, g.47657053_47657057del, g.47657064del, g.47657073dup,g.47657312_47676594del, g.47668611_47674615del, g.47672116_47675123del,g.47666463_47677632del, g.47666403_47677572del,g.(47657081_47672686)_(47710367_?)del,g.(47657081_47672686)_(47710367_?)inv,g.47671507_47675022delinsCATTCTCTTTGAAAA, g.47657278_47676557del,g.47672687_47672796del, g.(47657081_47672686)_(47672797_47690169)del,g.(47657081_47672686)_(47672797_47690169)del,g.(47657081_47672686)_(47693948_47698103)del,g.(47657081_47672686)_(47698202_47702163)del,g.(47657081_47672686)_(47708011_47709917)del, g.47672691dup,g.47672697dup, g.47672721_47672744delins47672748_47672771inv,g.47672728_47672729del, g.47672731dup, g.47672750_47672751insGG,g.47672755_47672758del, g.47672762_47672763del, g.47630466_47630494del,g.47686194_47697740dell, g.(47672797_47690169)_(47710367_?)del,g.(47672797_47690169)_(47690294_47693796)del,g.(47672797_47690169)_(47693948_47698103)del, g.47690170_47693947del,g.(47672797_47690169)_(47693948_47698103)del,g.(47672797_47690169)_(47693948_47698103)dup,g.(47672797_47690169)_(47705659_47707834)del, g.47690173del,g.47690191del, g.47690216_47690217dup, g.47690227del, g.47690227dup,g.47690228_47690232del, g.47690230_47690231del, g.47690240del,g.47690240_47690243del, g.47630475del, g.47630475_47630476del,g.47690259_47690260delinsCT, g.47690277dup, g.47690280del,g.47690283dup, g.(47690294_47693796)_(47702410_47703505)del,g.47630484_47630485insG, g.47693838_47693839del, g.47693862del,g.47693864del, g.47693873del, g.47693880dup, g.47693913del,g.47693924_47693925dup, g.47630493del, g.47697730_47706125del,g.(47693948_47698103)_(47710367_?)del,g.(47693948_47698103)_(47698202_47702163)del,g.(47693948_47698103)_(47705659_47707834)del, g.47698107del,g.47698109del, g.47698109_47698110insA, g.47630496del, g.47698118del,g.47698125del, g.47698129dup, g.47698138_47698139del,g.47698142_47698146del, g.47698144dup, g.47698147_47698148del,g.47698147_47698148dup, g.47698147_47698148insT, g.47698159del,g.47698162del, g.47698506_47703472del, g.47701803_47708848del,g.(47698202_47702163)_(47710367_?)del,g.(47698202_47702163)_(47702410_47703505)del,g.(47698202_47702163)_(47703711_47705410)del,g.(47698202_47702163)_(47705659_47707834)del, g.47702164del,g.47702175_47702176insA, g.47702183_47702186del,g.47702185_47702186insCT, g.47702190_47702192del, g.47702191dup,g.47702192_47702193del, g.47702213del, g.47702231del, g.47702242dup,g.47702257del, g.47702262_47702263dup, g.47630516_47630517dup,g.47630517del, g.47630517dup, g.47702289_47702290inv,g.47702293_47702296del, g.47702301dup, g.47702315del, g.47702315del,g.47702328_47702329del, g.47630522dup, g.47702339del,g.47702371_47702374dup, g.47702384_47702385del, g.47702386_47702389del,g.47702388del, g.47702388_47702389del, g.47702390del,g.47702390_47702391del, g.47702400_47702401del, g.47703506_47703710del,g.47703506_47708010del, g.47703510del, g.47703515del,g.47703521_47703522del, g.47703535_47703536del, g.47703546_47703547del,g.47703548_47703611dup, g.47630534del, g.47703571dup,g.47703574_47703581del, g.47703585dup, g.47630350del,g.47632107_47668733del, g.47703613del,g.(47630542_47635539)_(47643569_47656880)del,g.(47630542_47635539)_(47643569_47656880)inv,g.(47630542_47635539)_(47657081_47672686)del, g.47635540_47657080del,g.(47630542_47635539)_(47672797_47690169)del,g.(47630542_47635539)_(47690294_47693796)del,g.(47630542_47635539)_(47705659_47707834)del, g.47635540_47635694del,g.(47630542_47635539)_(47635695_47637232)del,g.(47630542_47635539)_(47635695_47637232)del,g.(47630542_47635539)_(47637512_47639552)del, g.47703635dup,g.47703641dup, g.47635542_47635549del, g.47703660_47703663del,g.47703667dup, g.47630351dup, g.47703704del, g.47703826_47707938del,g.(47703711_47705410)_(47705659_47707834)del, g.47705428_47705431del,g.47705437_47705438insA, g.47635551_47635552del, g.47705440_47705441del,g.47705461del, g.47705490del, g.47705494del, g.47705495del,g.47635557_47635558del, g.47705505del, g.47705535dup, g.47705547del,g.47705560_47705561dup, g.47705561dup, g.47705562dup, g.47705588del,g.47705608_47705609del, g.47705618dup, g.47705627dup,g.47635571_47635601delins(217), g.(47705659_47707834)_(47710367_?)del,g.(47705659_47707834)_(47708011_47709917)del, g.47707842_47707843del,g.47707861del, g.47707861_47707874dup, g.47707878_47707884del,g.47707878_47707884del, g.47707883del, g.47707895_47707905dell,g.47707897del, g.47707901_47707902del, g.47707905_47707906del,g.47707921del, g.47635583dup, g.47635583_47635584del,g.47707969_47707973del, g.47707996_47707997ins(115),g.47708009_47708010del, g.(47708011_47709917)_(47710367_?)del,g.47635591_47635592del, g.47635597_47635618dup, g.47635606_47635607del,g.47630359dup, g.47635672del, g.47635675_47635678del, g.47630364dup,g.47635680dup, g.47636862_47639040del, g.47636781_47638831del,g.47636753_47638155del, g.47636552_47638597del,g.(47635695_47637232)_(47643569_47656880)del,g.(47635695_47637232)_(47643569_47656880)del,g.(47635695_47637232)_(47657081_47672686)del,g.(47635695_47637232)_(47672797_47690169)del,g.(47635695_47637232)_(47698202_47702163)del,g.(47635695_47637232)_(47637512_47639552)del,g.(47635695_47637232)_(47641558_47643434)del, g.47637234del,g.47637246_47637247del, g.47637253_47637254dell, g.47637254_47637255del,g.47637254_47637255del, g.47637265del, g.47637274del, g.47637282del,g.47637320del, g.47637372_47637375del, g.47637377_47637449dup,g.47637379del, g.47637384del, g.47637394_47637395del,g.47637396_47637397del, g.47637417del, g.47637427_47637435del,g.47637437_47637439del, g.47637453del, g.47637458dup,g.47637479_47637482dup, g.47637482dup, g.47637504_47637505del,g.47637508_47637511del, g.47638050_47653430del, g.47638302_47648462del,g.47638478_47648643dell, g.(47637512_47639552)_(47710367_?)del,g.(47637512_47639552)_(47643569_47656880)del, g.47639553_47643568del,g.(47637512_47639552)_(47657081_47672686)del,g.(47637512_47639552)_(47657081_47672686)del,g.(47637512_47639552)_(47672797_47690169)del,g.(47637512_47639552)_(47639700_47641407)del,g.(47637512_47639552)_(47641558_47643434)del, g.47639557_47639561del,g.47639582_47639586delinsTAAT, g.47639583_47639584del, g.47639594del,g.47639594dup, g.47639598del, g.47639603_47639604del,g.47639611_47639612del, g.47639612del, g.47639618_47639621del,g.47639624_47639628delinsTTA, g.47630401dup, g.47639632dup,g.47639638_47639641dup, g.47639638_47639641dup, g.47639639del,g.47639639del, g.47639642dup, g.47630403_47630404insC, g.47639653del,g.47639666del, g.47639666_47639669del, g.47639668del,g.47639670_47639673delinsTT, g.47639674_47639675dup,g.47639695_47639696del, g.47639707_47642985del, g.47641402_47642007del,g.(47639700_47641407)_(47643569_47656880)del, g.47641408_47643568del,g.(47639700_47641407)_(47657081_47672686)del,g.(47639700_47641407)_(47672797_47690169)del,g.(47639700_47641407)_(47641558_47643434)del,g.(47639700_47641407)_(47641558_47643434)del, g.47641410del,g.47641425_47641426del, g.47641426_47641429del, g.47630412del,g.47641451del, g.47641454dup, g.47641455dup, g.47641469del,g.47641478del, g.47641488_47641491del, g.47641496_47641497del,g.47641503del, g.47641513_47641514dup, g.47641530_47641537dup,g.47642509_47655432del, g.(47641558_47643434)_(47643569_47656880)del,g.(47641558_47643434)_(47693948_47698103)del, g.47630424_47630433del,g.47643450dup, g.47643462_47643463del, g.47643462_47643463ins(4),g.47643464_47643465insNC_000022.10:35788169_35788352, g.47643465dup.

MutS Homolog 3 (MSH3) is a human homologue of the bacterial mismatchrepair protein MutS that participates in the mismatch repair (MMR)system. MSH3 typically forms the heterodimer MutSβ with MSH2 in order tocorrect long insertion/deletion loops and base-base mispairs inmicrosatellites during DNA synthesis. Deficient capacity for MMR isfound in approximately 15% of colorectal cancers, and somatic mutationsin the MSH3 gene can be found in nearly 50% of MMR-deficient colorectalcancers. Examples of the mutations in MSH3 include, but are not limitedto, g.79970809del.

MSH6 encodes MutS homologue 6 (MSH6), a member of the Mutator S (MutS)family of proteins that are involved in DNA mismatch repair (MMR). TheMSH6 protein forms a heterodimer with MutS homologue 2 (MSH2) in bothhuman and yeast. Human MSH2/6 recognizes single base-base mismatches andshort insertion/deletion loops. Upon recognition of a mismatch, MSH2/6complex binds and exchanges ADP for ATP, resulting in a conformationalchange to the complex that precedes base pair dissolution, baseexcision, and repair.

MSH6 mutations include frameshift and/or nonsense mutations and canresult in non-functional MSH6 and loss of protein expression. Examplesinclude a frameshift mutation at MSH6 amino acid residue 290 and acompounding missense T1189I.

Inactivating MSH6 mutations can be detected in cancers by routinediagnostics methods. These methods include, but are not limited to,obtaining cancer cells and other diagnostic indicators such asperipheral blood mononuclear cells (PBMCs), PBMC subpopulations,circulating blasts (CD34+ cells), circulating tumor cells andcirculating exosomescancer cells by biopsy and blood tests and byobtaining lymphatic or other bodily fluids. It is then determined fromthe cancer cells or other diagnostic indicators whether the cancerexhibits an inactivating MSH6 mutation is by methodology known in theart, for example, direct DNA sequencing and multiplex ligation dependentprobe amplification, RNA sequencing (RNA-Seq), microarray, quantitativePCR, or NanoString™ gene expression panels, or MSH6 protein byimmunohistochemistry, flow cytometry, immunocytochemistry or Westernblot. Methods for identifying inactivating MSH6 mutations are disclosedin Houlleberghs H, Goverde A, Lusseveld J, Dekker M, Bruno M J, et al.(2017) Suspected Lynch syndrome associated MSH6 variants: A functionalassay to determine their pathogenicity. PLOS Genetics 13(5):

e1006765. https://doi.org/10.1371/journal.pgen.1006765.

Examples of the mutations in MSH6 include, but are not limited to,g.48032846_48032849del, g.48032846_48032849del, g.48032846_48032849del,g.48033337_48033342del, g.48033420_48033422del,g.(?_48010221)_(48034092)del, g.(?_48010221)_(48018263_48023032)del,g.47998510_48020183del, g.48007276_48020272del, g.48026207del,g.48026223del, g.48026223del, g.48026257_48026261del,g.48026261_48026265del, g.48026312_48026313del, g.48026398del,g.48026543_48026544dup, g.48026693dup, g.48026702del, g.48026712del,g.48026718dup, g.48026736_48026737delinsAG, g.48026736_48026737delinsG,g.48026750_4802675 del, g.48026754_48026757del, g.48026756_48026759del,g.48026759_48026760del, g.48026906del, g.48026928_48026931del,g.48026941dup, g.48026991del, g.48027023_48027024del, g.48027079del,g.48027079_48027082dup, g.48027167_48027168del, g.48027172_48027173dup,g.48027178_48027185del, g.48027184_48027185del, g.48027272_48027275del,g.48027470_48027471del, g.48027501_48027502del,g.48027501_48027502delTG, g.48027657dup, g.48027691_48027694del,g.48027733_48027736dup, g.48027794_48027796delinsC,g.48027841_48027842del, g.48027887del, g.48027890dup,g.48027973_48027980del, g.48028067del, g.48028098del, g.48028106del,g.48028175_48028176del, g.48028241_48028242del,g.48028241_48028242delTT, g.48028272_48028284dup,g.48028277_48028278del, g.48030558_48030559del, g.48030126_48032394del,g.48030568del, g.48030581_48030584del, g.48030584_48030585dup,g.48030607del, g.48030645_48030646insT, g.48030647del, g.48030647dup,g.48030649dup, g.48030654_48030660del, g.48030659dup,g.48030697_48030698del, g.48030698del, g.48030706del, g.48030710dup,g.48030727_48030728insC, g.48030765_48030829del,c.3438+797_3438+798insTATins1839_3439-428,c.3438+797_3438+798insTATins1839_3439-428, g.48032121_48032122del,g.48032123_48032124del, g.48032124dup, g.48032126_48032129del,g.48032129_48032130insA, g.48032129_48032132dup,g.(48032167_48032756)_(48034092_?)del, g.48032809_48032812del,g.48032835dup, g.48032846_48032849del, g.48033374_48033402dup,g.48033395_48033398del, g.48033421_48033433del, g.48033425_48033428dup,g.48033453_48033454insA, g.48033494_48033523del, g.48033495_48033496del,g.48033593dup, g.48033610_48033613dup, g.48033629_48033635del,g.48033636_48033639dup, g.48033676_48033682del, g.48033707dup,g.48033709_48033716dup, g.48033721_48033724dup, g.48033727_48033730dup,g.48033728_48033746dup,g.(48033742_48033743)_(48033742_48033743)ins(32), g.48033746dup,g.48033748_48033751del, g.48033758_48033768del,g.48033773_48033774insATCA, g.48033773_48033776dup,g.48033785_48033789dup, g.48033887_48033910inv,g.(48018263_48023032)_(48032167_48032756)del,g.(48018263_48023032)_(48023203_48025749)del, g.48023097_48023098del,g.48025773dup, g.48025832del, g.48025860_48025861insT,g.48025884_48025885del, g.48025967dup.

MutL homolog 1, colon cancer, nonpolyposis type 2 (E. coli) is a proteinthat in humans is encoded by the MLH1 gene located on Chromosome 3. Itis a gene commonly associated with hereditary nonpolyposis colorectalcancer.

Examples of the mutations in MSH6 include, but are not limited to,g.37089113_37089115del, g.37089175del, g.37090379_37090393del,g.37038201_37038202del, g.37042531_37042542del, g.37053339_37053355del,g.37053354del, g.37053590_37053591insT, g.37034841_37092337del,g.(?_37034841)_(37092337_?)del, g.(?_37034841)_(37061955_37067127)del,g.(?_37034841)_(37035155_37038109)del,g.(?_37034841)_(37035155_37038109)del,g.(?_37034841)_(37070424_37081676)del,g.(?_37034841)_(37083823_37089009)del, g.37034841_37083822del,g.(?_37034841)_(37038201_37042445)del,g.(?_37034841)_(37042545_37045891)del, g.37034841_37042544del,g.(?_37034841)_(37042545_37045891)del,g.(?_37034841)_(37042545_37045891)del,g.(?_37034841)_(37045966_37048481)del,g.(?_37034841)_(37050397_37053310)del,g.(?_37034841)_(37059091_37061800)del, g.37034658_37038806del,g.36961079_37138741del, g.37061923del, g.37061927del, g.37061933del,g.37061939del, g.37061942dup, g.37035140_37035141del, g.37070417del,g.37070417_37070418insT, g.37070419dup, g.37070422_37070423insT,g.37080355_37083368del, g.(37070424_37081676)_(37092337_?)del,g.(37070424_37081676)_(37081786_37083758)del,g.(37070424_37081676)_(37083823_37089009)del, g.37038148_37038151del,g.37038149del, g.37038149dup, g.37081690_37081691del,g.37081691_37081692del, g.37081706_37081708del, g.37081710_37081711del,g.37035053_37035066del, g.37038154del, g.37038154_37038157del,g.37081738_37081739del, g.37081740del, g.37081753dup,g.37081757_37081761dup, g.37081782_37081783insAAGT,g.37081787_37081793delinsATTT,g.(37081786_37083758)_(37083823_37089009)del,g.(37081786_37083758)_(37089175_37090007)del, g.37083759del,g.37083780dup, g.37083781_37083784del, g.37083781_37083784delCTCA,g.37083808_37083809del, g.37083816del, g.37086069_37089606del,g.37084092_37089247del, g.37084590_37089786del,g.(37083823_37089009)_(37092337_?)del,g.(37083823_37089009)_(37089175_37090007)del, g.37089010_37089174del,g.(37083823_37089009)_(37090509_37091976)del, g.37089023del,g.37089026_37089027del, g.37089027del, g.37089036del, g.37089036dup,g.37038168dup, g.37089042del, g.37089047del, g.37089050_37089053del,g.37089056_37089057dell, g.37089061_37089062del, g.37089078_37089096del,g.37089090dup, g.37089099dup, g.37089107_37089110dup,g.37089109_37089110del, g.37089130_37089132del,g.37089130_37089132delAAG, g.37089131delinsTTCTT, g.37089133del,g.37089133delG, g.37089144del, g.37089155del, g.37089155_37089161del,g.37089158_37089161del, g.37089162_37089166del, g.37089171del,g.(37089175_37090007)_(37090101_37090394)del, g.37035056_37035072del,g.37090013del, g.37090015dup, g.37038183_37038184del,g.37090024_37090037dup, g.37090025_37090053dup, g.37090027dup,g.37038184dup, g.37090031_37090032insT, g.37090041del, g.37090057del,g.37090064_37090067del, g.37038188del, g.37090082del,g.37090086_37090087del, g.37090087_37090088del,g.37090097_37090101delinsC, g.37090099del, g.37038191dup,g.(37090101_37090394)_(37092337_?)del, g.37035057_37035073del,g.37090405dup, g.37090411_37090415del, g.37090414del, g.37038194del,g.37038198del, g.37090472_37090478del, g.37039445_37059613dup,g.37039760_37052440del, g.37090481_37090482del, g.37090483_37090484del,g.37090483_37092045del, g.37040732_37043185delinsACATAGTA,g.37042445_37042446del, g.(37038201_37042445)_(37042545_37045891)del,g.(37038201_37042445)_(37048555_37050304)del,g.(37038201_37042445)_(37050397_37053310)del,g.(37038201_37042445)_(37053591_37055922)del, g.37090497_37090498del,g.37090497_37090498delTC, g.37090504_37090507del,g.(37090509_37091976)_(37092337_?)del,g.(37090509_37091976)_(37092337_?)dup, g.37091977_37091978del,g.37091978_37091987del, g.37042448_37042451del, g.37091984_37091990del,g.37042451_37042453del, g.37092020_37092021del, g.37092022_37092068dup,g.37092027_37092028del, g.37092027_37092028dup, g.37092030dup,g.37092052_37092055del, g.37092054_37092055del, g.37092068_37092071dup,g.37092091dup, g.37092094_37092097delins(30), g.37092096_37092106del,g.37092097del, g.37092125_37092126delAA, g.37092125_37092126del,g.37092139_37092142dup, g.37092142dup, g.37035060dup,g.37042469_37042470del, g.37042470del, g.37042482dup, g.37042485del,g.37042499del, g.37042546dup, g.37044472_37046589del,g.37045648_37049941del, g.37045095_37054651del, g.37045072_37046861del,g.(37042545_37045891)_(37045966_37048481)del,g.(37042545_37045891)_(37092337_?)del,g.(37042545_37045891)_(37048555_37050304)del,g.(37042545_37045891)_(37050397_37053310)del, g.37045892_37050396del,g.37035069del, g.37045926del, g.37045931del, g.37045939_37045940dup,g.37045957_37045958del, g.37045963del, g.37035075del,g.37048067_37049287del, g.(37045966_37048481)_(37048555_37050304)del,g.(37045966_37048481)_(37050397_37053310)del, g.37048483del,g.37048483_37048503delinsT, g.37048486_37048487delinsGTT, g.37048489del,g.37048490del, g.37035076_37035077insCCCA, g.37035077_37035078dup,g.37048505_37048508del, g.37048521del, g.37048529dup, g.37035082dup,g.37049873_37052281del, g.37049839_37052249del, g.37049800_37052209del,g.37049640_37050445del, g.37050305_37050396del,g.(37048555_37050304)_(37050397_37053310)del, g.37050305_37050396del,g.37050319_37050320del, g.37050339del, g.37050348del,g.37050353_37050354del, g.37050354dup, g.37050364del,g.37050375_37050376insGA, g.37035090del, g.37050382_37050383delinsAT,g.37050382_37050383delinsCT, g.37050390_37050396del,g.37052950_37060990del, g.(37050397_37053310)_(37067499_37070274)dup,g.(37050397_37053310)_(37053591_37055922)del,g.(37050397_37053310)_(37056036_37058996)del, g.37053353del,g.37053510_37053511del, g.37035099del, g.37053545_37053546insT,g.37053562del, g.37053578del, g.37053578dup, g.37053585del,g.37053586_37053589del, g.37053591del, g.37053590_37053591delinsAT,g.37055920_37055921del, g.37055914_37055938del,g.(37053591_37055922)_(37070424_37081676)del,g.(37053591_37055922)_(37083823_37089009)del,g.(37053591_37055922)_(37059091_37061800)del, g.37035105del,g.37055928dup, g.37035106_37035116del, g.37055938del, g.37035108del,g.37055972_37055975del, g.37055976_37055979del, g.37035111del,g.37055990dup, g.37035114del, g.37035116del, g.37056036del,g.37056037dup, g.37058993_37059001del,g.(37056036_37058996)_(37070424_37081676)del,g.(37056036_37058996)_(37059091_37061800)del, g.37058997_37059000del,g.37059014_37059017del, g.37059017_37059021del, g.37059027_37059030dup,g.37035122del, g.37059062_37059063insT, g.37059065_37059066del,g.37059066del, g.37059066dup, g.37059072_37059073del,g.37059072_37059073dup, g.37059090_37059093del, g.37061595_37061913del,g.37061308_37066756del, g.37061207_37063077del,g.(37059091_37061800)_(37092337_?)del,g.(37059091_37061800)_(37061955_37067127)del, g.37061801_37061954del,g.(37059091_37061800)_(37083823_37089009)del, g.37061803dup,g.37061804del, g.37061817del, g.37061837_37061838dup, g.37061844del,g.37061851dup, g.37061855dup, g.37061870del, g.37061904_37061906del,g.37061910del, g.37035047del, g.[37049179_37051317delinsTG;37051667_37054327delinsCA].

Human PMS2 related genes are located at bands 7p12, 7p13, 7q11, and7q22. Exons 1 through 5 of these homologues share high degree ofidentity to human PMS2. The product of this gene is involved in DNAmismatch repair. The protein forms a heterodimer with MLH1 and thiscomplex interacts with MSH2 bound to mismatched bases. Defects in thisgene are associated with hereditary nonpolyposis colorectal cancer, withTurcot syndrome, and are a cause of supratentorial primitiveneuroectodermal tumors.

Examples of the mutations in PMS2 include, but are not limited to,g.(?_6012870)_(6048737_?)del, g.6012870_6048737del,g.(6027252_6029430)_(6048737_?)del, g.(6045663_6048627)_(6048737_?)del,g.6029554del, g.6029499dup, g.6029495_6029496de,g.6029462_6029463delinsTAAA, g.5992485_6028601del,g.(6018328_6022454)_(6027252_6029430)del,g.(6013174_6017218)_(6027252_6029430)del, g.6027226_6027227ins(20),g.6027175del, g.6027090dup, g.6036705_6044207delinsCG, g.6026666dup,g.6026628del, g.6043671del, g.6026565dup, g.6026565dupT,g.6018315_6018316del, g.6018306_6018310del, g.6018306_6018310delAGTTA,g.6043633_6043634dup, g.6018256_6018259del, g.6015623_6017501del,g.6016429_6017479del, g.6017300_6017303del, g.6045579_6045674delinsATTT,g.(6043690_6045522)_(6045663_6048627)del,g.(?_6012870)_(6042268_6043320)del,g.(6035265_6036956)_(6042268_6043320)del, g.6038283_6039384del,g.6038901del, g.6038851dup, g.(6035265_6036956)_(6037055_6038738)del,g.6037019_6037024delinsCTTCACACACA, g.6036980del, g.6036958dup,g.6035323_6035324insJN866832.1,g.(6022623_6026389)_(6035265_6036956)del,g.(6031689_6035164)_(6035265_6036956)del, g.6035204_6035207del,g.6035205_6035206del, g.(?_6012870)_(6031689_6035164)del,g.(6027252_6029430)_(6031689_6035164)del,g.(6029587_6031603)_(6031689_6035164)del, g.6028725_6029882del,g.(?_6012870)_(6029587_6031603)del.

The present invention provides a method of treating patients with Lynchsyndrome to reduce the likelihood of from developing or treating cancersderived from Lynch syndrome, by administering to the subject aneffective amount of one or more disclosed compounds, or apharmaceutically acceptable salt thereof, or the correspondingpharmaceutical composition.

Lynch syndrome is a hereditary disorder caused by a mutation in amismatch repair gene in which affected individuals have a higher thannormal chance of developing colorectal cancer, endometrial cancer, andvarious other types of aggressive cancers, often at a young age—alsocalled hereditary nonpolyposis colon cancer (HNPCC).

The mutations of specific mismatch repair (MMR) genes including but notlimited to MLH1, MSH2, MSH6, PMS2, and EPCAM-TACSTD1 deletions areresponsible for Lynch syndrome. These genes work in repairing mistakesmade when DNA is copied in preparation for cell division. The defects inthe genes disallow repair of DNA mistakes and as cells divide, errorsstack and uncontrollable cell growth may result in cancer.

Those with Lynch syndrome carry up to an 85% risk of contracting coloncancer as well as a higher than average risk for endometrial cancer,stomach cancer, pancreatic cancer, kidney/ureter tract cancer,hepatobiliary tract cancer, gastric tract cancer, prostate cancer,ovarian cancer, gallbladder duct cancer, brain cancer, small intestinecancer, breast cancer, and skin cancer.

Thus, in one embodiment for the disclosed method, the method is a methodof treating cancer derived from Lynch syndrome, selected from the groupconsisting of colon cancer, endometrial cancer, stomach cancer,pancreatic cancer, kidney/ureter tract cancer, hepatobiliary tractcancer, gastric tract cancer, prostate cancer, ovarian cancer,gallbladder duct cancer, brain cancer, small intestine cancer, breastcancer, and skin cancer.

In yet another embodiment, the method is a method of treating autoimmunedisease. Exemplary autoimmune diseases include lupus erythematosus;Wiskott-Aldrich syndrome; autoimmune lymphoproliferative syndrome;myasthenia gravis; rheumatoid arthritis (RA); lupus nephritis; multiplesclerosis; systemic lupus erythematosis; discoid lupus; subacutecutaneous lupus erythematosus; cutaneous lupus erythematosus includingchilblain lupus erythematosus; chronic arthritis; Sjogren's syndrome;inflammatory chronic rhinosinusitis; colitis; celiac disease;inflammatory bowel disease; Barrett's esophagus; inflammatory gastritis;autoimmune nephritis; autoimmune vasculitis; autoimmune hepatitis;autoimmune carditis; autoimmune encephalitis; autoimmune diabetes;autoimmune diabetes nephritis; psoriasis; Graft-versus-host disease(GvHD); and autoimmune mediated hematological disease.

In one aspect of this embodiment, the method is a method of treatingimmune deficiency selected from the group consisting of AutoimmuneLymphoproliferative Syndrome (ALPS), Autoimmune polyglandular syndrometype 1 (APS-1), BENTA Disease, Caspase Eight Deficiency State (CEDS),Chronic Granulomatous Disease (CGD), Common Variable Immunodeficiency(CVID), Congenital Neutropenia Syndromes, CTLA4 Deficiency, DOCK8Deficiency, GATA2 Deficiency, Glycosylation Disorders WithImmunodeficiency, hyper-immunoglobulin E syndrome (HIES),Hyper-Immunoglobulin M (Hyper-IgM) Syndromes, Leukocyte adhesiondeficiency (LAD), LRBA deficiency, PI3 Kinase disease, PLCG2-associatedantibody deficiency and immune dysregulation (PLAID), severe combinedimmunodeficiency (SCID), STAT3 gain-of-function disease, Warts,Hypogammaglobulinemia, Infections, and Myelokathexis Syndrome (WHIMS),X-Linked Agammaglobulinemia (XLA), X-Linked Lymphoproliferative Disease(XLP), and XMEN Disease.

As used herein, the term “immune deficiency” refers to a condition inwhich a portion or some portions of cell components constituting animmune system are defective or dysfunction, so that a normal immunemechanism is damaged. In other words, “immune deficiency” means acondition under which: congenital immunity and/or acquired immunity aresuppressed and/or decreased. In some embodiments, the immune-deficiencysubject is an immunocompromised subject. Non-limiting examples of immunedeficiencies can include AIDS, hypogammaglobulinemia,agammaglobulinemia, granulocyte deficiency, chronic granulomatousdisease, asplenia, SCID, complement deficiency, and/or sickle cellanemia.

In another aspect of this embodiment, the method is a method of treatinga neurodegenerative disorder selected from the group consisting ofmultiple sclerosis, Parkinson's disease (PD), Alzheimer's disease (AD),Dentatorubropallidoluysian atrophy (DRPLA), Huntington's Disease (HD),Spinocerebellar ataxia Type 1 (SCA1), Spinocerebellar ataxia Type 2(SCA2), Spinocerebellar ataxia Type 3 (SCA3), Spinocerebellar ataxia 6(SCA6), Spinocerebellar ataxia Type 7 (SCA7), Spinocerebellar ataxiaType 8 (SCA8), Spinocerebellar ataxia Type 12 (SCA12), Spinocerebellarataxia Type 17 (SCA17), Spinobulbar Muscular Ataxia/Kennedy Disease(SBMA), Fargile X syndrome (FRAXA), Fragile XE mental retardation(FRAXE), and Myotonic dystrophy (DM).

A “subject” is a mammal, preferably a human, but can also be an animalin need of veterinary treatment, e.g., companion animals (e.g., dogs,cats, and the like), farm animals (e.g., cows, sheep, pigs, horses, andthe like) and laboratory animals (e.g., rats, mice, guinea pigs, and thelike).

In certain embodiments, the methods disclosed herein further compriseco-administering an effective amount of a DNA repair inhibitor, a DNAdamage response (DDR) inhibitor, a DNA damaging agent or animmunomodulatory agent to the subject being treated for cancer, inaddition to an effective amount of a disclosed RAD51 inhibitor.

The term “DNA repair inhibitor” refers to any agent that targetscomponents/processes which a cell uses to repair mutations or changes inDNA and restore the DNA to its original state and prevents the repair ofDNA. Examples of DNA repair inhibitors include: RPA inhibitors, APE1inhibitors, DNA ligase inhibitors, DNA polymerase inhibitors, Parpinhibitors etc.

The term “DNA damage response inhibitor” refers to any agent thattargets components/processes involved in detecting DNA lesions,signaling the presence of DNA damage, and/or promote the repair of DNAdamage. Examples of DNA damage response inhibitors include checkpointinhibitors, ATM and ATR inhibitiors, DNA-PK inhibitors, etc.

The term “DNA damaging agent” refers to any agent that directly orindirectly damages DNA for which homologous recombination could repairthe damage. The DNA damaging agents is selected from the groupconsisting of: exposure to a DNA damaging chemical; exposure to achemotherapeutic agent; exposure to a radiochemotherapy, and exposure toionizing or ultraviolet radiation. Specific examples of DNA-damagingchemotherapeutic agents include alkylating agents, nitrosoureas,anti-metabolites, plant alkaloids, plant extracts and radioisotopes.Specific examples of the chemotherapeutic agents also includeDNA-damaging drugs, for example, 5-fluorouracil (5-FU), capecitabine,S-1 (Tegafur, 5-chloro-2,4-dihydroxypyridine and oxonic acid),5-ethynyluracil, arabinosyl cytosine (ara-C), 5-azacytidine (5-AC),2′,2′-difluoro-2′-deoxycytidine (dFdC), purine antimetabolites(mercaptopurine, azathiopurine, thioguanine), gemcitabine hydrochlorine(Gemzar), pentostatin, allopurinol, 2-fluoro-arabinosyl-adenine(2F-ara-A), hydroxyurea, sulfur mustard (bischloroetyhylsulfide),mechlorethamine, melphalan, chlorambucil, cyclophosphamide, ifosfamide,thiotepa, AZQ, mitomycin C, dianhydrogalactitol, dibromoducitol, alkylsulfonate (busulfan), nitrosoureas (BCNU, CCNU, 4-methyl CCNU or ACNU),procarbazine, decarbazine, rebeccamycin, anthracyclins such asdoxorubicin (adriamycin; ADR), daunorubicin (Cerubicine), idarubicin(Idamycin) and epirubicin (Ellence), anthracyclin analogs such asmitoxantrone, actinimycin D, non-intercalating topoisomerase inhibitorssuch as epipodophyllotoxins (etoposide or VP16, teniposide or VM-26),podophylotoxin, bleomycin (Bleo), pepleomycin, compounds that formadducts with nucleic acid including platinum derivatives, e.g.,cisplatin (CDDP), trans analog of cisplatin, carboplatin, iproplatin,tetraplatin and oxaliplatin, as well as camptothecin, topotecan,irinotecan (CPT-11), and SN-38. Specific examples of nucleic aciddamaging treatments include radiation e.g., ultraviolet (UV), infrared(IR), or .alpha.-, .beta.-, or .gamma.-radiation, as well asenvironmental shock, e.g., hyperthermia.

“Immunomodulatory agent” means an agent that modulates an immuneresponse to an antigen but is not the antigen or derived from theantigen. “Modulate”, as used herein, refers to inducing, enhancing,suppressing, directing, or redirecting an immune response. Such agentsinclude immunostimulatory agents, such as adjuvants, that stimulate (orboost) an immune response to an antigen but is not an antigen or derivedfrom an antigen. There are several distinct types of immunomodulatoryagents, which include, but are not limited to, Toll-like Receptor (TLR)agonists and Toll-like Receptor (TLR) antagonists. Such agents alsoinclude immunosuppressants. The immunomodulatory agent is selected fromthe group consisting of immune checkpoint modulators, Toll-like receptor(TLR) agonists, cell-based therapies, cytokines and cancer vaccines.

In certain embodiments, the subject is determined to have an increasedlevel and/or activity of a DNA damage process or DNA editing enzyme. Inone aspect of this embodiment, the DNA editing enzyme is selected fromthe group consisting of activation induced cytidine deaminase (AID orAICDA), APOBEC2, APOBEC3A, APOBEC3C, APOBEC3D, APOBEC3F, APOBEC3G,APOBEC3H, APOBEC4, a Type 1 Topoisomerase, a Type 2 Topoisomerase,Recombination Activating Gene 1 (RAG 1), and Recombination ActivatingGene 2 (RAG2).

In certain embodiments, blood cells obtained from the subject have beendetermined to have a detectable level of activation-induced cytidinedeaminase (AID).

In certain embodiments, B cells obtained from the subject have beendetermined to have a detectable level of activation-induced cytidinedeaminase (AID).

In certain embodiments, the detectable level of activation-inducedcytidine deaminase (AID) is statistically significantly higher than thelevel of AID expressed in unactivated B-cells or normal non-immune cellsfrom a healthy subject.

Methods of Administration and Dosage Forms

The precise amount of compound administered to provide an “effectiveamount” to the subject will depend on the mode of administration, thetype, and severity of the disease, and on the characteristics of thesubject, such as general health, age, sex, body weight, and tolerance todrugs. The skilled artisan will be able to determine appropriate dosagesdepending on these and other factors. When administered in combinationwith other therapeutic agents, e.g., when administered in combinationwith an anti-cancer agent, an “effective amount” of any additionaltherapeutic agent(s) will depend on the type of drug used. Suitabledosages are known for approved therapeutic agents and can be adjusted bythe skilled artisan according to the condition of the subject, the typeof condition(s) being treated and the amount of a compound of theinvention being used by following, for example, dosages reported in theliterature and recommended in the Physician's Desk Reference (57th ed.,2003).

The term “effective amount” means an amount when administered to thesubject which results in beneficial or desired results, includingclinical results, e.g., inhibits, suppresses or reduces the symptoms ofthe condition being treated in the subject as compared to a control. Forexample, a therapeutically effective amount can be given in unit dosageform (e.g., 0.1 mg to about 50 g per day, alternatively from 1 mg toabout 5 grams per day).

The terms “administer”, “administering”, “administration”, and the like,as used herein, refer to methods that may be used to enable delivery ofcompositions to the desired site of biological action. These methodsinclude, but are not limited to, intraarticular (in the joints),intravenous, intramuscular, intratumoral, intradermal, intraperitoneal,subcutaneous, orally, topically, intrathecally, inhalationally,transdermally, rectally, and the like. Administration techniques thatcan be employed with the agents and methods described herein are foundin e.g., Goodman and Gilman, The Pharmacological Basis of Therapeutics,current ed.; Pergamon; and Remington's, Pharmaceutical Sciences (currentedition), Mack Publishing Co., Easton, Pa.

In addition, the disclosed RAD51 inhibitors can be co-administered withother therapeutic agents. As used herein, the terms “co-administration”,“administered in combination with”, and their grammatical equivalents,are meant to encompass administration of two or more therapeutic agentsto a single subject, and are intended to include treatment regimens inwhich the agents are administered by the same or different route ofadministration or at the same or different times. In some embodimentsthe one or more compounds described herein will be co-administered withother agents. These terms encompass administration of two or more agentsto the subject so that both agents and/or their metabolites are presentin the subject at the same time. They include simultaneousadministration in separate compositions, administration at differenttimes in separate compositions, and/or administration in a compositionin which both agents are present. Thus, in some embodiments, thecompounds described herein and the other agent(s) are administered in asingle composition. In some embodiments, the compounds described hereinand the other agent(s) are admixed in the composition.

The particular mode of administration and the dosage regimen will beselected by the attending clinician, taking into account the particularsof the case (e.g., the subject, the disease, the disease state involved,the particular treatment). Treatment can involve daily or multi-daily orless than daily (such as weekly or monthly etc.) doses over a period ofa few days to months, or even years. However, a person of ordinary skillin the art would immediately recognize appropriate and/or equivalentdoses looking at dosages of approved compositions for treating a a RAD51mediated disease using the disclosed RAD51 inhibitors for guidance.

The compounds or the corresponding pharmaceutical compositions taughtherein can be administered to a patient in a variety of forms dependingon the selected route of administration, as will be understood by thoseskilled in the art. The compounds of the present teachings may beadministered, for example, by oral, parenteral, buccal, sublingual,nasal, rectal, patch, pump or transdermal administration and thepharmaceutical compositions formulated accordingly. Parenteraladministration includes intravenous, intraperitoneal, subcutaneous,intramuscular, transepithelial, nasal, intrapulmonary, intrathecal,rectal and topical modes of administration. Parenteral administrationcan be by continuous infusion over a selected period of time.

The pharmaceutical composition of the invention is formulated to becompatible with its intended route of administration. In an embodiment,the composition is formulated in accordance with routine procedures as apharmaceutical composition adapted for intravenous, subcutaneous,intramuscular, oral, intranasal, or topical administration to humanbeings. In preferred embodiments, the pharmaceutical composition isformulated for intravenous administration.

Typically, for oral therapeutic administration, a compound of thepresent teachings may be incorporated with excipient and used in theform of ingestible tablets, buccal tablets, troches, capsules, elixirs,suspensions, syrups, wafers, and the like.

Typically for parenteral administration, solutions of a compound of thepresent teachings can generally be prepared in water suitably mixed witha surfactant such as hydroxypropylcellulose. Dispersions can also beprepared in glycerol, liquid polyethylene glycols, DMSO and mixturesthereof with or without alcohol, and in oils. Under ordinary conditionsof storage and use, these preparations contain a preservative to preventthe growth of microorganisms.

Typically, for injectable use, sterile aqueous solutions or dispersionof, and sterile powders of, a compound described herein for theextemporaneous preparation of sterile injectable solutions ordispersions are appropriate.

EXEMPLIFICATION Abbreviations

-   Ac acetyl-   ACN acetonitrile-   aq aqueous-   Bn benzyl-   Boc tert-butoxycarbonyl-   br. broad-   d doublet (only when used within 1H NMR spectra)-   DCM dichloromethane-   DIEA(DIPEA) diisopropylethylamine-   DMA dimethylacetamide-   DMAP 4-dimethylaminopyridine-   DMF N,N-dimethylformamide-   DMSO dimethylsulfoxide-   dppf 1,1′-bis(diphenylphosphino) ferrocene-   eq equivalent-   EtOAc ethyl acetate-   hr hour-   HBTU N,N,N′,N′,-tetramethyl-O-(1H-benzotriazol-1-yl)uronium    hexafluorophosphate-   HPLC high performance liquid chromatography-   LC-MS liquid chromatography coupled to mass spectrometry-   m multiplet-   MS ESI mass spectra, electrospray ionization-   NBS N-bromosuccinimide-   NMR nuclear magnetic resonance-   prep preparative-   Py pyridine-   s singlet-   sat saturated-   SFC supercritical fluid chromatography-   t triplet-   TEA triethylamine-   TFA trifluoroacetic acid-   THF tetrahydrofuran-   TLC thin layer chromatography-   Tol toluene

General Method Reaction Name A Substitution Reaction B Suzuki Reaction AC Deprotection of Boc group A (TFA) D Acylation Reaction E UreaFormation F Deprotection of Boc group B (HCl) G Reduction with Fe HCarbamate Formation I Hydrogenation J Bromination K Suzuki Reaction B LThionation M Cyclization N Coupling Reaction O Hydrolysis Reaction

Example 1. Synthesis of (S)-(1-methylpyrrolidin-2-yl)methyl(4-(2-(4-((benzylcarbamoyl)oxy)piperidin-1-yl)thiazol-5-yl)-3-(N-(tert-butyl)sulfamoyl)phenyl)carbamate

General Method D for Preparation of Sulfonamide Compound 12.

To a solution of5-amino-2-(2-bromothiazol-5-yl)-N-tert-butyl-benzenesulfonamide (5 g,12.8 mmol, 1 eq.) in DCM (30 mL) were added DMAP (156 mg, 1.3 mmol, 0.1eq.) and Ac₂O (1.96 g, 19.2 mmol, 1.5 eq.). The mixture was stirred at20° C. for 1 hr, and then washed with 1M HCl (50 mL) and sat.aq.Na₂CO₃(50 mL). The organic layer was dried over Na₂SO₄, filtered andconcentrated. The residue was purified by column chromatography (SiO₂,Petroleum ether:Ethyl acetate=5:1 to 2:1) to giveN-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl)phenyl]acetamide (1.5g, 3.5 mmol, 27% yield) as a yellow solid. ESI [M+H]=433.9/431.9

General Method A for Preparation of Sulfonamide Compound 13.

To a solution ofN-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl)phenyl]acetamide (700mg, 1.6 mmol, 1 eq.) in DMF (20 mL) were added K₂CO₃ (448 mg, 3.3 mmol,2 eq.) and piperidin-4-ol (246 mg, 2.4 mmol, 1.5 eq.). The mixture wasstirred at 100° C. for 12 hrs and then poured into H₂O (100 mL). Theaqueous phase was extracted with EtOAc (50 mL×3), the combined organiclayers were washed with brine (100 mL), dried over Na₂SO₄, filtered andconcentrated. The residue was purified by column chromatography (SiO₂,Petroleum ether:Ethyl acetate=1:2) to giveN-[3-(tert-butylsulfamoyl)-4-[2-(4-hydroxy-1-piperidyl)thiazol-5-yl]phenyl]acetamide(550 mg, 1.2 mmol, 13.2% yield) as a yellow solid. ¹H NMR (400 MHz,CHLOROFORM-d) δ=8.26 (dd, J=2.2, 8.6 Hz, 2H), 7.93 (d, J=2.4 Hz, 1H),7.37 (d, J=8.4 Hz, 1H), 7.31 (s, 1H), 4.02-3.94 (m, 1H), 3.88-3.80 (m,2H), 3.30 (ddd, J=3.5, 9.2, 13.1 Hz, 2H), 2.21 (s, 3H), 2.03-1.95 (m,2H), 1.67 (dtd, J=4.0, 8.7, 13.0 Hz, 2H), 1.09 (s, 9H). ESI [M+H]=453.1

General Method F for Preparation of Sulfonamide Compound 15.

[1-[5-[4-acetamido-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-4-piperidyl]N-benzylcarbamate

(380 mg, 649 umol, 1 eq.) was dissolved into HCl/MeOH (4 M, 20 mL) andthe mixture was stirred at 20° C. for 1 hr. The mixture wasconcentrated, diluted with sat.aq.Na₂CO₃ (20 mL) and extracted withEtOAc (20 mL×3). The combined organic layers were washed with brine (20mL), dried over Na₂SO₄, filtered and concentrated. The residue waspurified by column chromatography (SiO₂, Petroleum ether:Ethylacetate=10:1 to 1:1) to give[1-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-4-piperidyl]N-benzylcarbamate (300 mg, 552 umol, 85% yield) as a yellow solid. ESI[M+H]=544.2

General Method H for Preparation of Example 1.

To a solution of [(2S)-1-methylpyrrolidin-2-yl]methanol (42 mg, 367umol, 2 eq.) and DIEA (71.11 mg, 550 umol, 3 eq.) in MeCN (5 mL) wasadded a solution of[1-[5-[2-(tert-butylsulfamoyl)-4-[(4-nitrophenoxy)carbonylamino]phenyl]thiazol-2-yl]-4-piperidyl]N-benzylcarbamate (130 mg, 184 umol, 1 eq.) in DCM (2 mL) and themixture was refluxed for 1 hr. The mixture was concentrated and theresidue was purified by prep-HPLC to give(S)-(1-methylpyrrolidin-2-yl)methyl(4-(2-(4-((benzylcarbamoyl)oxy)piperidin-1-yl)thiazol-5-yl)-3-(N-(tert-butyl)sulfamoyl)phenyl)carbamate(19.87 mg, 28.8 umol, 15.7% yield, 99.3% purity) as a pale yellow solid.¹H NMR (400 MHz, DMSO-d6) δ=10.06 (s, 1H), 8.26 (d, J=2.0 Hz, 1H), 7.71(t, J=6.2 Hz, 1H), 7.60 (dd, J=2.3, 8.5 Hz, 1H), 7.37-7.14 (m, 7H), 6.94(s, 1H), 4.81-4.71 (m, 1H), 4.16 (br d, J=6.2 Hz, 2H), 4.10-4.03 (m,1H), 4.02-3.96 (m, 1H), 3.73-3.63 (m, 2H), 3.28 (br s, 2H), 2.94-2.87(m, 1H), 2.44-2.37 (m, 1H), 2.30 (s, 3H), 2.18-2.05 (m, 1H), 1.99-1.82(m, 3H), 1.68-1.52 (m, 5H), 1.12-1.03 (m, 9H). ESI [M+H]=685.2

Example 2. Synthesis of[1-[5-[2-(tert-butylsulfamoyl)-4-[[(2R)-1-methylpyrrolidin-2-yl]methoxycarbonylamino]phenyl]thiazol-2-yl]-4-piperidyl]N-benzylcarbamate

The following compound was synthesized via same method by the keyintermediate 16.

[1-[5-[2-(tert-butylsulfamoyl)-4-[[(2R)-1-methylpyrrolidin-2-yl]methoxycarbonylamino]phenyl]thiazol-2-yl]-4-piperidyl]N-benzylcarbamate

¹H NMR (400 MHz, DMSO-d6) δ=10.07 (s, 1H), 8.29 (d, J=2.2 Hz, 1H), 7.73(t, J=6.2 Hz, 1H), 7.63 (dd, J=2.3, 8.5 Hz, 1H), 7.39-7.19 (m, 7H), 6.94(s, 1H), 4.80 (td, J=4.1, 8.0 Hz, 1H), 4.20 (br d, J=6.1 Hz, 2H),4.14-4.07 (m, 1H), 4.06-3.99 (m, 1H), 3.72 (br d, J=13.6 Hz, 2H),3.29-3.22 (m, 2H), 2.98-2.92 (m, 1H), 2.45-2.41 (m, 1H), 2.33 (s, 3H),2.22-2.13 (m, 1H), 2.02-1.79 (m, 3H), 1.72-1.56 (m, 5H), 1.20-1.03 (m,9H). ESI [M+H]=685.2

Example 3. Synthesis of[1-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-4-piperidyl] N-isopropylcarbamate

Preparation of Compound 23.

General method A,1-benzyl-3-[3-(tert-butylsulfamoyl)-4-[2-(4-hydroxy-1-piperidyl)thiazol-5-yl]phenyl]urea. ESI [M+H]=544.1

Preparation of Ex. 3.

To a solution of1-benzyl-3-[3-(tert-butylsulfamoyl)-4-[2-(4-hydroxy-1-piperidyl)thiazol-5-yl]phenyl]urea (50 mg, 91.96 umol, 1 eq.) in Tol. (2 mL), wereadded DMAP (22.47 mg, 183.92 umol, 2 eq.) and[isopropyl(methyl)-azanylidene] methanone (78.26 mg, 919.62 umol, 10eq.). The mixture was stirred at 100° C. for 4 hrs and thenconcentrated. The residue was purified by prep-TLC (Petroleumether/EtOAc=1:3) and then acidic prep-HPLC to give[1-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-4-piperidyl]N-isopropylcarbamate (28.31 mg, 99.2% purity) as a yellow solid. ¹H NMR(400 MHz, METHANOL-d4) δ=8.29 (d, J=2.0 Hz, 1H), 7.71 (dd, J=2.0, 8.3Hz, 1H), 7.43 (d, J=8.8 Hz, 1H), 7.35 (d, J=3.9 Hz, 5H), 7.27 (dt,J=2.7, 5.7 Hz, 1H), 4.98-4.93 (m, 1H), 4.43 (s, 2H), 3.90-3.59 (m, 5H),2.19-2.07 (m, 2H), 1.99-1.86 (m, 2H), 1.22 (s, 9H), 1.16 (d, J=6.8 Hz,6H). ESI [M+H]=629.1

Example 4. Synthesis of Isopropyl4-[5-[2-(tert-butylsulfamoyl)-4-(2-pyridylmethylcarbamoylamino)phenyl]thiazol-2-yl]piperidine-1-carboxylate

Preparation of Compound 45

General method D, 4-nitrophenyl(4-bromo-3-(N-(tert-butyl)sulfamoyl)phenyl) carbamate. ESI [M+H]=474.1

General Method E for Preparation of Sulfonamide Compound 46.

To a solution of 2-pyridylmethanamine (352 mg, 3.3 mmol, 5 eq.) and DIEA(84 mg, 650 umol, 1 eq.) in DCM (3 mL) was added the solution of(4-nitrophenyl)N-[4-bromo-3-(tert-butylsulfamoyl)phenyl]carbamate (307mg, 650 umol, 1 eq.) in DCM (3 mL). The mixture was stirred at 25° C.for 1 hr, then diluted with DCM (30 mL) and washed with H₂O (20 mL×2).The organic layer was concentrated and the residue was purified byprep-TLC (SiO₂, Petroleum ether:Ethyl acetate=0:1) to give1-[4-bromo-3-(tert-butylsulfamoyl) phenyl]-3-(2-pyridylmethyl)urea (180mg, crude) as a yellow solid. ESI [M+H]=441.2/443.2

Preparation of Compound 47

A mixture of1-[4-bromo-3-(tert-butylsulfamoyl)phenyl]-3-(2-pyridylmethyl)urea (140mg, 317.21 umol, 1 eq.),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(322.21 mg, 1.27 mmol, 4 eq.), Pd(dppf)Cl₂ (116.05 mg, 158.61 umol, 0.5eq.) and KOAc (93.39 mg, 951.64 umol, 3 eq.) in dioxane (4 mL) wasdegassed and purged with N₂ for 3 times, and then the mixture wasstirred at 80° C. for 12 hrs under N₂ atmosphere. The mixture wasconcentrated and the residue was purified by column chromatography(SiO₂, Petroleum ether/Ethyl acetate=1/0 to 1:1) to give1-[3-(tert-butylsulfamoyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]-3-(2-pyridylmethyl)urea(50 mg, 89.37 umol, 28.17% yield, 87.3% purity). ESI [M+H]=489.4

General Method K for Preparation of Compound Ex. 4

A mixture of1-[3-(tert-butylsulfamoyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]-3-(2-pyridylmethyl)urea(26.38 mg, 54.01 umol, 1.2 eq.), isopropyl4-(5-bromothiazol-2-yl)piperidine-1-carboxylate (15 mg, 45.01 umol, 1eq.), Na₂CO₃ (9.54 mg, 90.02 umol, 2 eq.), KF (7.85 mg, 135.04 umol, 3eq.) and Pd(PPh₃)₄(5.20 mg, 4.50 umol, 0.1 eq.) in H₂O (0.1 mL)/EtOH(0.3 mL)/Tol. (0.3 mL) was degassed and purged with N₂ for 3 times, andthen the mixture was stirred at 85° C. for 12 hrs under N₂ atmosphere.The reaction mixture was filtered and concentrated. The residue waspurified by prep-TLC (SiO₂, Petroleum ether/Ethyl acetate=1/2) and thenprep-HPLC (TFA condition) to give isopropyl4-[5-[2-(tert-butylsulfamoyl)-4-(2-pyridylmethylcarbamoylamino)phenyl]thiazol-2-yl]piperidine-1-carboxylate(6.36 mg, 8.21 umol, 18.24% yield, 94.07% purity, TFA) as a pale yellowsolid. ¹H NMR (400 MHz, METHANOL-d4) δ=8.75 (br d, J=5.6 Hz, 1H), 8.53(br t, J=7.9 Hz, 1H), 8.34 (d, J=2.0 Hz, 1H), 8.04 (br d, J=8.1 Hz, 1H),7.92 (br t, J=6.5 Hz, 1H), 7.75 (s, 1H), 7.69 (dd, J=2.3, 8.4 Hz, 1H),7.39 (d, J=8.3 Hz, 1H), 4.94-4.92 (m, 1H), 4.77 (s, 2H), 4.24 (br d,J=13.4 Hz, 2H), 3.31-3.25 (m, 1H), 3.02 (br s, 2H), 2.15 (br d, J=11.5Hz, 2H), 1.76 (dq, J=4.2, 12.3 Hz, 2H), 1.28 (d, J=6.2 Hz, 6H), 1.11 (s,9H). ESI [M+H]=615.2

Example 5. Synthesis of Isopropyl4-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxycarbonylamino)phenyl]thiazol-2-yl]piperazine-1-carboxylate

Preparation of Compound 56

General method A, tert-butyl4-[5-[4-acetamido-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]piperazine-1-carboxylate. ESI [M+H]=538.2

General Method C for Preparation of Sulfonamide Compound 57.

To a solution of tert-butyl4-[5-[4-acetamido-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]piperazine-1-carboxylate(110 mg, 205 umol, 1 eq.) in DCM (2 mL) was added TFA (1 mL) and themixture was stirred at 25° C. for 30 mins. The mixture was concentratedto giveN-[3-(tert-butylsulfamoyl)-4-(2-piperazin-1-ylthiazol-5-yl)phenyl]acetamide(80 mg, 183 umol, 89.4% yield) as a yellow solid. ESI [M+H]=438.2

Preparation of Compound 58

General method D, isopropyl4-[5-[4-acetamido-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]piperazine-1-carboxylate. ESI [M+H]=524.1

Preparation of Compound 59

General method F, isopropyl4-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]piperazine-1-carboxylate.ESI [M+H]=482.1

Preparation of Compound Ex. 5

General method D, isopropyl4-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxycarbonylamino)phenyl]thiazol-2-yl]piperazine-1-carboxylate. ¹H NMR (400 MHz,METHANOL-d4) δ=8.22 (d, J=2.1 Hz, 1H), 7.56 (dd, J=2.2, 8.3 Hz, 1H),7.30 (d, J=8.3 Hz, 1H), 7.20 (s, 1H), 4.94-4.81 (m, 2H), 3.61-3.43 (m,8H), 1.20 (dd, J=6.2, 14.5 Hz, 12H), 1.07 (s, 9H). ESI [M+H]=568.3

Example 6. Synthesis of Isopropyl4-[5-[4-(benzyloxycarbonylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]piperazine-1-carboxylate

Scheme 5:

Preparation of Compound Ex. 6

General method D, isopropyl4-[5-[4-(benzyloxycarbonylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]piperazine-1-carboxylate.¹H NMR (400 MHz, METHANOL-d4) δ=8.33 (d, J=2.0 Hz, 1H), 7.68 (dd, J=2.2,8.4 Hz, 1H), 7.45-7.27 (m, 7H), 5.21 (s, 2H), 4.94-4.88 (m, 1H),3.68-3.51 (m, 8H), 1.27 (d, J=6.4 Hz, 6H), 1.19-1.11 (m, 9H). ESI[M+H]=616.3

Example 7. Synthesis of trans-isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(cyclopentoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate

General Method B for Preparation of Sulfonamide Compound 83.

To a mixture of tert-butyl(trans-4-(5-bromothiazol-2-yl)cyclohexyl)carbamate (1.3 g, 3.6 mmol, 2eq.) and isopropylN-[3-(tert-butylsulfamoyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]carbamate(800 mg, 1.8 mmol, 1 eq.) in dioxane (12 mL) and H₂O (2 mL) were addedNa₂CO₃ (579 mg, 5.5 mmol, 3 eq.) and Pd(dppf)Cl₂ (133 mg, 182 umol, 0.1eq.). The mixture was stirred at 80° C. for 12 hrs under N₂ atmosphereand then concentrated. The residue was diluted with H₂O (5 mL) andextracted with ethyl acetate (5 mL×3), dried over Na₂SO₄, filtered andconcentrated. The residue was purified by column chromatography (SiO₂,Petroleum ether:Ethyl acetate=1:0 to 3:1) to givetrans-isopropyl-N-[4-[2-[4-(tert-butoxycarbonylamino)cyclohexyl]thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]carbamate (660 mg, crude),in which 60 mg was purified by prep-HPLC (column: Agela Durashell C18150×25 5u; mobile phase: [water(0.04% NH3H2O)-ACN]; B %: 60%-90%, 10min) to give pure compound 83 (40.92 mg, 99.64% purity) as a white solidfor delivery. ¹H NMR (400 MHz, DMSO-d6) δ=10.06 (s, 1H), 8.32 (d, J=2.1Hz, 1H), 7.70-7.60 (m, 2H), 7.38 (d, J=8.4 Hz, 1H), 6.96 (s, 1H), 6.81(br d, J=7.9 Hz, 1H), 4.99-4.87 (m, 1H), 3.27 (br s, 1H), 2.94-2.84 (m,1H), 2.13 (br d, J=11.7 Hz, 2H), 1.90 (br d, J=11.1 Hz, 2H), 1.63-1.49(m, 2H), 1.39 (s, 9H), 1.33 (br d, J=14.4 Hz, 2H), 1.28 (d, J=6.4 Hz,6H), 1.07 (s, 9H). ESI [M+H]=595.3

Preparation of Compound 84

General method F, trans-isopropylN-[4-[2-(4-aminocyclohexyl)thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]carbamate.ESI [M+H]=495.2

Preparation of Ex. 7

General method D, trans-isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(cyclopentoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate.¹H NMR (400 MHz, DMSO-d6) δ=10.04 (s, 1H), 8.29 (d, J=2.2 Hz, 1H),7.61-7.59 (m, 2H), 7.35 (d, J=8.8 Hz, 1H), 7.00 (br d, J=7.9 Hz, 1H),6.96-6.93 (m, 1H), 4.96-4.87 (m, 2H), 3.29-3.22 (m, 1H), 2.87 (br t,J=11.8 Hz, 1H), 2.10 (br d, J=12.7 Hz, 2H), 1.88 (br d, J=12.7 Hz, 2H),1.75 (br s, 2H), 1.61-1.48 (m, 8H), 1.36-1.27 (m, 2H), 1.25 (d, J=6.1Hz, 6H), 1.04 (s, 9H). ESI [M+Na]=629.3

Example 8. Synthesis of [(2R)-1-methylpyrrolidin-2-yl]methylN-[4-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxycarbonylamino)phenyl]thiazol-2-yl]cyclohexyl]carbamate

Preparation of Compound 85

General method D,trans-(4-nitrophenyl)N-[4-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxycarbonylamino)phenyl]thiazol-2-yl]cyclohexyl]carbamate.ESI [M+H]=660.2

Preparation of Compound Ex. 8

General method H, [(2R)-1-methylpyrrolidin-2-yl]methylN-[4-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxycarbonylamino)phenyl]thiazol-2-yl]cyclohexyl]carbamate.¹H NMR (400 MHz, METHANOL-d4) δ=8.38 (s, 1H), 7.80-7.73 (m, 1H), 7.69(dd, J=2.2, 8.4 Hz, 1H), 7.40 (d, J=8.3 Hz, 1H), 5.05-5.00 (m, 1H), 4.48(br dd, J=3.0, 12.8 Hz, 1H), 4.21 (br dd, J=7.2, 12.6 Hz, 1H), 3.78-3.66(m, 2H), 3.57-3.46 (m, 1H), 3.22 (td, J=8.3, 11.2 Hz, 1H), 3.07-3.01 (m,3H), 2.43-1.99 (m, 8H), 1.98-1.88 (m, 1H), 1.80-1.66 (m, 2H), 1.47 (q,J=12.5 Hz, 2H), 1.34 (d, J=6.2 Hz, 6H), 1.20-1.12 (m, 9H). ESI[M+H]=636.2

Example 9. Synthesis of Isopropyl(3-(N-(tert-butyl)sulfamoyl)-4-(2-((1S,4r)-4-(((((S)-1-methylpyrrolidin-2-yl)methoxy)carbonyl)amino)cyclohexyl)thiazol-5-yl)phenyl)carbamate

The following compound was synthesized via same method by the keyintermediate 85.

¹H NMR (METHANOL-d4, 400 MHz): δ=8.34 (d, J=2.2 Hz, 1H), 7.71 (s, 1H),7.66 (dd, J=8.5, 2.1 Hz, 1H), 7.36 (d, J=8.4 Hz, 1H), 4.93-5.04 (m, 1H),4.46 (dd, J=13.0, 2.9 Hz, 1H), 4.18 (dd, J=13.0, 7.1 Hz, 1H), 3.64-3.75(m, 2H), 3.42-3.58 (m, 1H), 3.12-3.25 (m, 1H), 3.00-3.06 (m, 3H),1.97-2.39 (m, 8H), 1.84-1.96 (m, 1H), 1.64-1.77 (m, 2H), 1.44 (q, J=12.6Hz, 2H), 1.31 (d, J=6.2 Hz, 6H), 1.13 (s, 9H). ESI [M+H]=636.3

Example 10. Synthesis of trans-tert-butylN-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate

General method B, trans-tert-butylN-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate.¹H NMR (400 MHz, METHANOL-d4) δ=8.24 (d, J=2.4 Hz, 1H), 7.72-7.66 (m,2H), 7.37-7.29 (m, 5H), 7.28-7.21 (m, 1H), 4.40 (s, 2H), 3.39 (br t,J=11.6 Hz, 1H), 2.98 (tt, J=3.4, 12.1 Hz, 1H), 2.25-2.17 (m, 2H), 2.04(br d, J=11.5 Hz, 2H), 1.67 (dq, J=2.9, 12.9 Hz, 2H), 1.44 (s, 9H),1.41-1.32 (m, 2H), 1.10 (s, 9H). ESI [M+H]=642.3

Example 11. Synthesis of [(2R)-1-methylpyrrolidin-2-yl]methylN-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate

Preparation of Compound 86

General method F,trans-1-[4-[2-(4-aminocyclohexyl)thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]-3-benzyl-urea.ESI [M+H]=542.3

Preparation of Compound 87

General method D,trans-(4-nitrophenyl)N-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate.

Preparation of Ex. 11

General method H, [(2R)-1-methylpyrrolidin-2-yl]methylN-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate.¹H NMR (400 MHz, METHANOL-d4) δ=8.30 (d, J=2.2 Hz, 1H), 7.79 (s, 1H),7.70 (dd, J=2.3, 8.4 Hz, 1H), 7.42-7.31 (m, 5H), 7.27 (dt, J=2.6, 5.7Hz, 1H), 4.54-4.41 (m, 3H), 4.21 (dd, J=7.1, 12.7 Hz, 1H), 3.79-3.66 (m,2H), 3.52 (br t, J=11.6 Hz, 1H), 3.26-3.17 (m, 1H), 3.04 (s, 3H),2.42-1.98 (m, 8H), 1.97-1.87 (m, 1H), 1.80-1.65 (m, 2H), 1.54-1.40 (m,2H), 1.15 (s, 9H). ESI [M+H]=683.3

Example 12. Synthesis of ((S)-1-methylpyrrolidin-2-yl)methyl((1r,4S)-4-(5-(4-(3-benzylureido)-2-(N-(tert-butyl)sulfamoyl)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 87.

1H NMR (400 MHz, METHANOL-d4) δ=8.29 (d, J=2.3 Hz, 1H), 7.83-7.74 (m,1H), 7.73-7.65 (m, 1H), 7.43-7.31 (m, 5H), 7.30-7.22 (m, 1H), 4.55-4.35(m, 3H), 4.21 (br dd, J=7.1, 12.7 Hz, 1H), 3.79-3.61 (m, 2H), 3.58-3.42(m, 1H), 3.23 (br s, 1H), 3.08-3.02 (m, 3H), 2.43-1.98 (m, 8H),1.97-1.86 (m, 1H), 1.80-1.65 (m, 2H), 1.54-1.39 (m, 2H), 1.19-1.07 (m,9H). ESI [M+H]=683.3

Example 13. Synthesis of Isopropyl(S)-(4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(1-phenylethyl)ureido)phenyl)thiazol-2-yl)bicyclo[2.2.2]octan-1-yl)carbamate

Preparation of Compound 89

General method D,4-(isopropoxycarbonylamino)bicyclo[2.2.2]octane-1-carboxylate.

General Method O for Preparation of Compound 90

To a solution of methyl4-(isopropoxycarbonylamino)bicyclo[2.2.2]octane-1-carboxylate (400 mg,1.49 mmol, 1 eq.) in MeOH (5 mL), was added LiOH (106.70 mg, 4.46 mmol,3 eq.) in H₂O (5 mL) and the mixture was stirred at 50° C. for 1 hr. Themixture was concentrated to remove MeOH and then extracted with MTBE (10mL*2). The pH of aqueous phase was adjusted to 1-2 by adding 4 N HClsolution and then extracted with EtOAc (10 mL*3). The combined organicphase was dried over Na₂SO₄, filtered and concentrated to give4-(isopropoxycarbonylamino)bicyclo[2.2.2]octane-1-carboxylic acid (270mg, crude) as a yellow solid. ¹H NMR (400 MHz, CHLOROFORM-d) δ=4.86 (brd, J=6.4 Hz, 1H), 4.46 (br s, 1H), 2.00-1.80 (m, 12H), 1.23 (br d, J=5.9Hz, 6H).

General Method N for Preparation of Compound 91

To a solution of4-(isopropoxycarbonylamino)bicyclo[2.2.2]octane-1-carboxylic acid (270mg, 1.06 mmol, 1 eq.) in DCM (5 mL), were added DMF (7.73 mg, 105.75umol, 0.1 eq.) and (COCl)₂ (201.34 mg, 1.59 mmol, 1.5 eq.). The mixturewas stirred at 26° C. for 0.2 hr and then concentrated. The residue wasdissolved into THF (5 mL) and was added into NH₃.H₂O (741.24 mg, 5.29mmol, 5 eq.) in dioxane (5 mL) dropwise. Then the mixture was stirred at26° C. for 0.3 hr. The mixture was concentrated and diluted with EtOAc(20 mL) and washed with H₂O (10 mL). The organic phase was dried overNa₂SO₄, filtered and concentrated to give isopropylN-(1-carbamoyl-4-bicyclo[2.2.2]octanyl)carbamate as a yellow solid. ¹HNMR (400 MHz, CHLOROFORM-d) δ=5.43 (br s, 1H), 5.17 (br s, 1H),4.85-4.71 (m, 1H), 4.35 (br s, 1H), 1.82 (s, 12H), 1.13 (d, J=6.2 Hz,6H). ESI[M+H]=255.2

General Method L for Preparation of Compound 92

To a solution of isopropylN-(1-carbamoyl-4-bicyclo[2.2.2]octanyl)carbamate (60 mg, 235.92 umol, 1eq.) in 2-Me-THF (2 mL), was added2,4-bis(4-methoxyphenyl)-2,4-dithioxo-1,3,2,4dithiadiphosphetane (95.42mg, 235.92 umol, 1 eq.) and the mixture was stirred at 80° C. for 0.5hr. The mixture was poured into sat.aq.Na₂CO₃ (10 mL) and extracted withEtOAc (10 mL*3). The combined organic phase was dried over Na₂SO₄,filtered and concentrated. The residue was purified by prep-TLC(Petroleum ether/Ethyl acetate=2:3) to give isopropylN-(1-carbamothioyl-4-bicyclo[2.2.2]octanyl)carbamate (30 mg, crude) as ayellow solid. ESI [M+H]=271.1

General Method M for Preparation of Compound 93

To a solution of isopropylN-(1-carbamothioyl-4-bicyclo[2.2.2]octanyl)carbamate (85 mg, 314.36umol, 1 eq.) in EtOH (2 mL), were added TsOH.H₂O (119.59 mg, 628.72umol, 2 eq.) and 2-bromo-1,1-diethoxy-ethane (123.90 mg, 628.72 umol, 2eq.). The mixture was stirred at 80° C. for 1 hr and then concentratedand diluted with EtOAc (30 mL). The mixture was washed withsat.aq.Na₂CO₃ (10 mL*2) and the combined organic phase was dried overNa₂SO₄, filtered and concentrated. The residue was purified by prep-TLC(Petroleum ether/Ethyl acetate=3:1) to give isopropylN-(1-thiazol-2-yl-4-bicyclo[2.2.2]octanyl)carbamate (80 mg, crude) as ayellow solid. ESI [M+H]=295.3

Preparation of Compound 94

General method J, isopropyl N-[1-(5-bromothiazol-2-yl)-4-bicyclo[2.2.2]octanyl]carbamate. ESI[M+H]=375.2/373.2

Preparation of Compound 95

General method B, isopropylN-[1-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-4-bicyclo[2.2.2]octanyl]carbamate. ESI [M+H]=521.2

Preparation of Compound 96

General method D,(4-nitrophenyl)N-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)-1-bicyclo[2.2.2]octanyl]thiazol-5-yl]phenyl]carbamate.ESI [M+H]=686.4

Preparation of Ex. 13

General method E, isopropylN-[1-[5-[2-(tert-butylsulfamoyl)-4-[[(1S)-1-phenylethyl]carbamoylamino]phenyl]thiazol-2-yl]-4-bicyclo[2.2.2]octanyl]carbamate.¹H NMR (400 MHz, METHANOL-d4) δ=8.21 (d, J=2.3 Hz, 1H), 7.75 (s, 1H),7.67 (dd, J=2.3, 8.3 Hz, 1H), 7.41-7.32 (m, 5H), 7.29-7.23 (m, 1H),4.99-4.93 (m, 1H), 4.83-4.76 (m, 1H), 2.17-2.08 (m, 6H), 2.07-1.99 (m,6H), 1.51 (d, J=7.0 Hz, 3H), 1.22 (br d, J=6.0 Hz, 6H), 1.12 (s, 9H).ESI [M+H]=668.3

Example 14. Synthesis of Isopropyl(R)-(4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(1-phenylethyl)ureido)phenyl)thiazol-2-yl)bicyclo[2.2.2]octan-1-yl)carbamate

The following compound was synthesized via same method by the keyintermediate 96.

¹H NMR (400 MHz, METHANOL-d4) δ=8.18 (d, J=2.6 Hz, 1H), 7.74-7.69 (m,1H), 7.64 (dd, J=2.2, 8.3 Hz, 1H), 7.38-7.29 (m, 5H), 7.26-7.19 (m, 1H),4.92 (q, J=7.0 Hz, 1H), 4.82-4.71 (m, 1H), 2.15-2.05 (m, 6H), 2.04-1.95(m, 6H), 1.48 (d, J=7.0 Hz, 3H), 1.20 (br d, J=6.1 Hz, 6H), 1.09 (s,9H). ESI [M+H]=668.3

Example 15. Synthesis of Isopropyl(S)-(4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(1-(pyridin-2-yl)ethyl)ureido)phenyl)thiazol-2-yl)bicyclo[2.2.2]octan-1-yl)carbamate

The following compound was synthesized via same method by the keyintermediate 96.

¹H NMR (400 MHz, METHANOL-d4) δ=8.72 (d, J=5.6 Hz, 1H), 8.46 (t, J=7.9Hz, 1H), 8.28 (d, J=2.3 Hz, 1H), 8.02 (d, J=7.9 Hz, 1H), 7.85 (t, J=6.6Hz, 1H), 7.72 (s, 1H), 7.63 (dd, J=2.3, 8.4 Hz, 1H), 7.35 (d, J=8.4 Hz,1H), 5.13 (q, J=7.1 Hz, 1H), 4.84-4.74 (m, 1H), 2.15-2.07 (m, 6H),2.07-1.98 (m, 6H), 1.65 (d, J=7.1 Hz, 3H), 1.22 (br d, J=6.1 Hz, 6H),1.09 (s, 9H). ESI [M+H]=669.2

Example 16. Synthesis of Isopropyl(R)-(4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(1-(pyridin-2-yl)ethyl)ureido)phenyl)thiazol-2-yl)bicyclo[2.2.2]octan-1-yl)carbamate

The following compound was synthesized via same method by the keyintermediate 96.

¹H NMR (400 MHz, METHANOL-d4) δ=8.75 (d, J=5.1 Hz, 1H), 8.57 (dt, J=1.5,7.9 Hz, 1H), 8.29 (d, J=2.2 Hz, 1H), 8.11 (d, J=8.1 Hz, 1H), 7.98-7.90(m, 1H), 7.75 (s, 1H), 7.62 (dd, J=2.3, 8.3 Hz, 1H), 7.36 (d, J=8.4 Hz,1H), 5.15 (q, J=7.1 Hz, 1H), 4.84-4.75 (m, 1H), 2.16-2.07 (m, 6H),2.06-1.97 (m, 6H), 1.67 (d, J=7.2 Hz, 3H), 1.22 (br d, J=6.1 Hz, 6H),1.09 (s, 9H). ESI [M+H]=669.2

Example 17. Synthesis of IsopropylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)-1-bicyclo[2.2.2]octanyl]thiazol-5-yl]phenyl]carbamate

Scheme 10:

Preparation of Compound Ex. 17

General method B, isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)-1-bicyclo[2.2.2]octanyl]thiazol-5-yl]thiazol-phenyl]carbamate. ¹HNMR (400 MHz, METHANOL-d4) δ=8.33 (d, J=2.0 Hz, 1H), 7.74 (s, 1H), 7.66(dd, J=2.2, 8.4 Hz, 1H), 7.36 (d, J=8.4 Hz, 1H), 5.03-4.91 (m, 1H), 4.78(br d, J=6.4 Hz, 1H), 2.16-2.05 (m, 6H), 2.04-1.95 (m, 6H), 1.31 (d,J=6.4 Hz, 6H), 1.20 (br d, J=6.0 Hz, 6H), 1.11 (s, 9H). ESI [M+H]=607.3

Example 18. Synthesis of Isopropyl(4-(5-(4-(3-benzylureido)-2-(N-(tert-butyl)sulfamoyl)phenyl)thiazol-2-yl)bicyclo[2.2.2]octan-1-yl)carbamate

The following compound was synthesized via same method by the keyintermediate 94.

¹H NMR (400 MHz, METHANOL-d4) δ=8.26 (d, J=2.2 Hz, 1H), 7.77 (s, 1H),7.72 (dd, J=2.4, 8.4 Hz, 1H), 7.38-7.33 (m, 5H), 7.27 (td, J=2.6, 8.6Hz, 1H), 4.80 (br d, J=5.7 Hz, 1H), 4.43 (s, 2H), 2.17-2.10 (m, 6H),2.06-2.00 (m, 6H), 1.25-1.20 (m, 6H), 1.13 (s, 9H). ESI [M+H]=654.1

Example 19. Synthesis of Isopropyl(4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(pyridin-2-ylmethyl)ureido)phenyl)thiazol-2-yl)bicyclo[2.2.2]octan-1-yl)carbamate

The following compound was synthesized via same method by the keyintermediate 94.

¹H NMR (400 MHz, METHANOL-d4) δ=8.73 (d, J=5.7 Hz, 1H), 8.51 (dt, J=1.3,7.9 Hz, 1H), 8.31 (d, J=2.2 Hz, 1H), 8.02 (d, J=8.3 Hz, 1H), 7.90 (t,J=6.8 Hz, 1H), 7.72 (s, 1H), 7.66 (dd, J=2.2, 8.3 Hz, 1H), 7.35 (d,J=8.3 Hz, 1H), 4.75 (s, 3H), 2.15-1.93 (m, 12H), 1.20 (br d, J=6.1 Hz,6H), 1.07 (s, 9H). ESI [M+H]=655.3

Example 20. Synthesis of trans-isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropylcarbamoyloxy)cyclohexyl]thiazol-5-yl]phenyl]carbamate

Preparation of Compound 111.

To a solution of trans-4-hydroxycyclohexanecarboxylic acid (1 g, 6.94mmol, 1 eq.) in MeCN (10 mL), were added HBTU (2.89 g, 7.63 mmol, 1.1eq.) and TEA (2.11 g, 20.81 mmol, 2.90 mL, 3 eq.) followed by additionof NH₄Cl (742.07 mg, 13.87 mmol, 2 eq.) and the mixture was stirred at26° C. for 1 hr. The mixture was then filtered and dried to givetrans-4-hydroxycyclohexanecarboxamide (1 g, crude) as a white solidwhich can be used without any purification.

Preparation of Compound 112.

To a solution of trans-4-hydroxycyclohexanecarboxamide (12 g, 83.81mmol, 1 eq.) in DCM (150 mL), were added IMIDAZOLE (11.41 g, 167.62mmol, 2 eq.) and tert-butyl-chloro-diphenyl-silane (27.64 g, 100.57mmol, 25.83 mL, 1.2 eq.). The mixture was stirred at 26° C. for 12 hrsand then poured into 1N HCl (200 mL) and the organic phase was washedwith sat.aq.Na₂CO₃ (100 mL). The organic phase was dried over Na₂SO₄,filtered and concentrated. The residue was washed with a solution(petroleum ether:EtOAc=10:1, 100 mL) and then filtered. The filter cakewas dried to givetrans4-[tert-butyl(diphenyl)silyl]oxycyclohexanecarboxamide (31.5 g,crude). ¹H NMR (400 MHz, METHANOL-d4) δ=7.72-7.64 (m, 4H), 7.49-7.36 (m,6H), 3.69-3.55 (m, 1H), 2.21-2.08 (m, 1H), 1.96-1.83 (m, 2H), 1.82-1.69(m, 2H), 1.50-1.23 (m, 4H), 1.12-0.98 (m, 9H).

Preparation of Compound 113.

General method L,trans-4-[tert-butyl(diphenyl)silyl]oxycyclohexanecarbothioamide. ¹H NMR(400 MHz, METHANOL-d4) δ=7.73-7.64 (m, 4H), 7.52-7.36 (m, 6H), 3.73-3.59(m, 1H), 2.59-2.44 (m, 1H), 1.91 (br d, J=8.3 Hz, 2H), 1.80-1.66 (m,2H), 1.57-1.35 (m, 4H), 1.08-1.04 (m, 9H). ESI [M+H]=398.1

Preparation of Compound 114.

General method M,trans-tert-butyl-diphenyl-(4-thiazol-2-ylcyclohexoxy)silane. ¹H NMR (400MHz, METHANOL-d4) δ=7.63-7.55 (m, 5H), 7.42-7.25 (m, 7H), 4.05-3.96 (m,1H), 2.97 (tt, J=3.7, 11.7 Hz, 1H), 2.05 (dq, J=3.2, 12.5 Hz, 2H),1.84-1.62 (m, 4H), 1.44 (tt, J=2.9, 13.5 Hz, 2H), 1.01-0.98 (m, 9H). ESI[M+H]=422.2

Preparation of Compound 114A.

General method J,trans-[4-(5-bromothiazol-2-yl)cyclohexoxy]-tert-butyl-diphenyl-silane(1.8 g, crude). ESI [M+H]=502.0

Preparation of Compound 115.

To a solution oftrans-[4-(5-bromothiazol-2-yl)cyclohexoxy]-tert-butyl-diphenyl-silane(1.5 g, 3.00 mmol, 1 eq.) in THF (10 mL), was added TBAF (1 M, 4.49 mL,1.5 eq.) and the mixture was stirred at 26° C. for 12 hrs and thenconcentrated. The residue was purified by silica gel chromatography(Petroleum ether/Ethyl acetate=20:1-1:1) to affordtrans-4-(5-bromothiazol-2-yl)cyclohexanol (500 mg, 1.91 mmol, 63.64%yield) as a yellow solid.

Preparation of Compound 116.

To a solution of trans-4-(5-bromothiazol-2-yl)cyclohexanol (500 mg, 1.91mmol, 1 eq.) in DMF (5 mL), were added 2-isocyanatopropane (486.93 mg,5.72 mmol, 3 eq.) and DIEA (739.47 mg, 5.72 mmol, 3 eq.). The mixturewas stirred at 100° C. for 40 hrs and then poured into H₂O (50 mL) andextracted with EtOAc (10 mL*3). The combined organic phase was driedover Na₂SO₄, filtered and concentrated. The residue was purified bysilica gel chromatography (Petroleum ether/Ethyl acetate=10:1-1:1) toafford trans-[4-(5-bromothiazol-2-yl)cyclohexyl] N-isopropylcarbamate(180 mg, 518.33 umol, 27.18% yield) as a yellow solid. ¹H NMR (400 MHz,METHANOL-d4) δ=7.61 (s, 1H), 3.85-3.64 (m, 2H), 3.15-2.99 (m, 1H),1.98-1.83 (m, 6H), 1.73 (br d, J=13.5 Hz, 2H), 1.14-1.12 (m, 6H). ESI[M+H]=347.1/349.1

Preparation of Ex. 20.

General method B, trans-isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropylcarbamoyloxy)cyclohexyl]thiazol-5-yl]phenyl]carbamate.¹H NMR (400 MHz, METHANOL-d4) δ=8.34 (d, J=2.0 Hz, 1H), 7.75 (s, 1H),7.67 (dd, J=2.0, 8.4 Hz, 1H), 7.37 (d, J=8.4 Hz, 1H), 5.05-4.93 (m, 1H),4.89 (br s, 1H), 3.83-3.64 (m, 1H), 3.15 (br s, 1H), 2.12-1.88 (m, 6H),1.84-1.64 (m, 2H), 1.31 (d, J=6.2 Hz, 6H), 1.19-1.04 (m, 15H). ESI[M+H]=581.4

Example 21. Synthesis of(1r,4r)-4-(5-(4-(3-benzylureido)-2-(N-(tert-butyl)sulfamoyl)phenyl)thiazol-2-yl)cyclohexyl isopropylcarbamate

The following compound was synthesized via same method by the keyintermediate 116.

¹H NMR (400 MHz, METHANOL-d4) δ=8.24 (d, J=2.2 Hz, 1H), 7.74 (s, 1H),7.69 (dd, J=2.4, 8.4 Hz, 1H), 7.40-7.28 (m, 5H), 7.28-7.17 (m, 1H), 4.89(br s, 1H), 4.41 (s, 2H), 3.79-3.64 (m, 1H), 3.14 (br s, 1H), 2.04-1.94(m, 6H), 1.84-1.65 (m, 2H), 1.16-1.08 (m, 15H). ESI [M+H]=628.4

Example 22. Synthesis of trans-isopropylN-[4-[5-[2-(tert-butylsulfamoyl)-4-(4-pyridylmethylcarbamoylamino)phenyl]thiazol-2-yl]cyclohexyl]carbamate

Preparation of Compound 117.

General method K, trans-isopropylN-[4-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate.1H NMR (400 MHz, METHANOL-d4) δ=7.68-7.57 (s, 1H), 7.43 (d, J=2.4 Hz,1H), 7.13 (d, J=8.2 Hz, 1H), 6.82 (dd, J=2.4, 8.2 Hz, 1H), 4.84-4.77 (m,1H), 3.44 (tt, J=3.9, 11.5 Hz, 1H), 2.97 (tt, J=3.6, 12.1 Hz, 1H),2.26-2.13 (m, 2H), 2.11-2.02 (m, 2H), 1.67 (dq, J=3.0, 12.9 Hz, 2H),1.49-1.32 (m, 2H), 1.22 (dd, J=2.5, 6.7 Hz, 6H), 1.09 (s, 9H). ESI[M+H]=495.2

Preparation of Compound 118.

General method D,trans-(4-nitrophenyl)N-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate.ESI [M+H]=660.1

Preparation of Ex. 22.

General method E, trans-isopropylN-[4-[5-[2-(tert-butylsulfamoyl)-4-(4-pyridylmethylcarbamoylamino)phenyl]thiazol-2-yl]cyclohexyl]carbamate.¹H NMR (400 MHz, METHANOL-d4) δ=8.79 (d, J=6.7 Hz, 2H), 8.34 (d, J=2.3Hz, 1H), 8.07 (d, J=6.6 Hz, 2H), 7.76 (s, 1H), 7.69 (dd, J=2.3, 8.4 Hz,1H), 7.39 (d, J=8.3 Hz, 1H), 4.85-4.83 (m, 1H), 4.73 (s, 2H), 3.47 (tt,J=3.6, 11.5 Hz, 1H), 3.04 (tt, J=3.4, 12.0 Hz, 1H), 2.30-2.19 (m, 2H),2.09 (br d, J=10.1 Hz, 2H), 1.71 (dq, J=3.0, 12.9 Hz, 2H), 1.43 (dq,J=3.3, 12.6 Hz, 2H), 1.24 (br d, J=6.1 Hz, 6H), 1.12 (s, 9H). ESI[M+H]=629.2

Example 23. Synthesis of Isopropyl((1r,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(4-fluorobenzyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.27 (d, J=2.3 Hz, 1H), 7.77 (s, 1H),7.71 (dd, J=2.4, 8.4 Hz, 1H), 7.38 (td, J=2.7, 8.6 Hz, 3H), 7.12-7.04(m, 2H), 5.00-4.92 (m, 1H), 4.41 (s, 2H), 3.54-3.41 (m, 1H), 3.05 (tt,J=3.5, 12.0 Hz, 1H),2.13-2.05 (m, 2H), 2.05-2.04 (m, 2H), 1.72 (dq,J=3.0, 12.8 Hz, 2H), 1.43 (dq, J=3.2, 12.5 Hz, 2H), 1.24 (br d, J=6.1Hz, 6H), 1.14 (s, 9H). ESI [M+H]=646.2

Example 24. Synthesis of Isopropyl((1R,4r)-4-(5-(2-(N-isopropylsulfamoyl)-4-(3-((R)-1-phenylethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.20 (d, J=2.2 Hz, 1H), 7.73 (s, 1H),7.64 (dd, J=2.4, 8.4 Hz, 1H), 7.38-7.29 (m, 5H), 7.26-7.19 (m, 1H),4.94-4.89 (m, 1H), 4.81 (br d, J=6.2 Hz, 1H), 3.44 (tt, J=3.9, 11.6 Hz,1H), 3.01 (tt, J=3.5, 12.0 Hz, 1H), 2.26-2.17 (m, 2H), 2.10-2.01 (m,2H), 1.68 (dq, J=2.9, 12.8 Hz, 2H), 1.48 (d, J=7.1 Hz, 3H), 1.43-1.33(m, 2H), 1.21 (br d, J=6.2 Hz, 6H), 1.10 (s, 9H). ESI [M+H]=642.3

Example 25. Synthesis of Isopropyl((1r,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(pyridin-3-ylmethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.84 (s, 1H), 8.74 (d, J=5.7 Hz, 1H),8.60 (d, J=8.2 Hz, 1H), 8.30 (d, J=2.2 Hz, 1H), 8.04 (dd, J=5.8, 7.8 Hz,1H), 7.72 (s, 1H), 7.65 (dd, J=2.4, 8.4 Hz, 1H), 7.36 (d, J=8.4 Hz, 1H),4.85-4.77 (m, 1H), 4.60 (s, 2H), 3.44 (tt, J=3.9, 11.5 Hz, 1H),3.10-2.93 (m, 1H), 2.28-2.17 (m, 2H), 2.13-2.01 (m, 2H), 1.69 (dq,J=3.0, 12.8 Hz, 2H), 1.50-1.33 (m, 2H), 1.22 (br d, J=6.2 Hz, 6H), 1.09(s, 9H). ESI [M+H]=629.3

Example 26. Synthesis of Isopropyl((1S,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((S)-1-phenylethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.19 (d, J=2.4 Hz, 1H), 7.70 (s, 1H),7.65 (dd, J=2.4, 8.4 Hz, 1H), 7.39-7.30 (m, 5H), 7.26-7.21 (m, 1H), 4.93(q, J=6.5 Hz, 1H), 4.83-4.81 (m, 1H), 3.49-3.40 (m, 1H), 3.05-2.96 (m,1H), 2.22 (br d, J=11.7 Hz, 2H), 2.06 (br d, J=11.5 Hz, 2H), 1.73-1.62(m, 2H), 1.49 (d, J=6.8 Hz, 3H), 1.43-1.36 (m, 2H), 1.22 (br d, J=6.0Hz, 6H), 1.10 (s, 9H). ESI[M+H]=642.3

Example 27. Synthesis of Isopropyl((1r,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(3-fluorobenzyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.27 (d, J=2.2 Hz, 1H), 7.74 (s, 1H),7.71 (dd, J=2.3, 8.3 Hz, 1H), 7.40-7.32 (m, 2H), 7.18 (d, J=7.7 Hz, 1H),7.10 (br d, J=9.9 Hz, 1H), 6.99 (dt, J=2.2, 8.5 Hz, 1H), 4.87 (br s,1H), 4.44 (s, 2H), 3.47 (tt, J=3.8, 11.6 Hz, 1H), 3.02 (tt, J=3.5, 12.0Hz, 1H), 2.24 (br d, J=12.2 Hz, 2H), 2.14-2.04 (m, 2H), 1.71 (dq, J=3.0,12.9 Hz, 2H), 1.50-1.36 (m, 2H), 1.24 (br d, J=6.1 Hz, 6H), 1.14 (s,9H). ESI [M+H]=646.2

Example 28. Synthesis of Isopropyl((1r,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(2-fluorobenzyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.23 (d, J=2.2 Hz, 1H), 7.71 (s, 1H),7.68 (dd, J=2.4, 8.4 Hz, 1H), 7.41 (dt, J=1.5, 7.6 Hz, 1H), 7.35 (d,J=8.4 Hz, 1H), 7.29 (ddt, J=1.8, 5.5, 7.7 Hz, 1H), 7.15 (dt, J=1.0, 7.6Hz, 1H), 7.11-7.05 (m, 1H), 4.81 (br s, 1H), 4.46 (s, 2H), 3.45 (tt,J=3.7, 11.6 Hz, 1H), 3.00 (tt, J=3.5, 12.0 Hz, 1H), 2.27-2.18 (m, 2H),2.06 (br d, J=10.1 Hz, 2H), 1.69 (dq, J=3.1, 12.9 Hz, 2H), 1.46-1.34 (m,2H), 1.22 (br d, J=6.2 Hz, 6H), 1.11 (s, 9H). ESI [M+H]=646.2

Example 29. Synthesis of Isopropyl((1r,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(pyridin-2-ylmethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.79-8.73 (m, 1H), 8.56 (dt, J=1.5, 7.9Hz, 1H), 8.35 (d, J=2.2 Hz, 1H), 8.06 (d, J=8.1 Hz, 1H), 7.94 (t, J=6.4Hz, 1H), 7.77 (s, 1H), 7.69 (dd, J=2.3, 8.4 Hz, 1H), 7.39 (d, J=8.3 Hz,1H), 4.86-4.83 (m, 1H), 4.79 (s, 2H), 3.47 (tt, J=3.9, 11.6 Hz, 1H),3.05 (tt, J=3.5, 12.0 Hz, 1H), 2.29-2.21 (m, 2H), 2.09 (br d, J=10.1 Hz,2H), 1.71 (dq, J=2.9, 12.8 Hz, 2H), 1.43 (dq, J=3.3, 12.6 Hz, 2H), 1.24(br d, J=6.2 Hz, 6H), 1.11 (s, 9H). ESI [M+H]=629.3

Example 30. Synthesis of Isopropyl((1R,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((R)-1-(pyridin-2-yl)ethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.73 (d, J=5.7 Hz, 1H), 8.53 (dt, J=1.8,7.9 Hz, 1H), 8.28 (d, J=2.2 Hz, 1H), 8.08 (d, J=8.3 Hz, 1H), 7.94-7.85(m, 1H), 7.80-7.68 (m, 1H), 7.61 (dd, J=2.2, 8.3 Hz, 1H), 7.35 (d, J=8.3Hz, 1H), 5.13 (q, J=7.0 Hz, 1H), 4.85-4.78 (m, 1H), 3.49-3.39 (m, 1H),3.08-2.96 (m, 1H), 2.22 (br d, J=12.3 Hz, 2H), 2.06 (br d, J=10.1 Hz,2H), 1.76-1.61 (m, 5H), 1.49-1.34 (m, 2H), 1.22 (br d, J=6.1 Hz, 6H),1.14-1.02 (m, 9H). ESI [M+H]=643.3

Example 31. Synthesis of Isopropyl((1r,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((3-fluoropyridin-2-yl)methyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.42 (d, J=4.8 Hz, 1H), 8.28 (d, J=2.2Hz, 1H), 7.78-7.62 (m, 3H), 7.44 (td, J=4.4, 8.5 Hz, 1H), 7.38 (d, J=8.4Hz, 1H), 4.86-4.81 (m, 1H), 4.65 (s, 2H), 3.47 (ddd, J=4.0, 7.7, 11.4Hz, 1H), 3.10-3.00 (m, 1H), 2.25 (br d, J=12.1 Hz, 2H), 2.09 (br d,J=10.5 Hz, 2H), 1.72 (dq, J=3.0, 12.9 Hz, 2H), 1.43 (dq, J=3.2, 12.6 Hz,2H), 1.24 (br d, J=6.1 Hz, 6H), 1.14 (s, 9H). ESI [M+H]=647.2

Example 32. Synthesis of Isopropyl((1S,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((S)-1-(pyridin-2-yl)ethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.72 (d, J=5.3 Hz, 1H), 8.50 (dt, J=1.8,7.9 Hz, 1H), 8.27 (d, J=2.2 Hz, 1H), 8.05 (d, J=8.3 Hz, 1H), 7.88 (t,J=6.6 Hz, 1H), 7.72 (s, 1H), 7.61 (dd, J=2.6, 8.3 Hz, 1H), 7.34 (d,J=8.8 Hz, 1H), 5.12 (q, J=7.0 Hz, 1H), 4.87-4.68 (m, 1H), 3.51-3.38 (m,1H), 3.01 (tt, J=3.3, 12.0 Hz, 1H), 2.21 (br d, J=11.8 Hz, 2H),2.10-1.97 (m, 2H), 1.76-1.57 (m, 5H), 1.48-1.33 (m, 2H), 1.22 (d, J=6.1Hz, 6H), 1.13-1.00 (m, 9H). ESI [M+H]=643.3

Example 33. Synthesis of trans-4-piperidylmethylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate

Preparation of Compound 119.

General method H, trans-tert-butyl4-[[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamoyloxymethyl]piperidine-1-carboxylate.ESI [M+H]=736.5

Preparation of Ex. 33.

General method C, trans-4-piperidylmethylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate.¹H NMR (400 MHz, METHANOL-d4) δ=8.37 (d, J=2.2 Hz, 1H), 7.78 (s, 1H),7.68 (br d, J=7.9 Hz, 1H), 7.40 (d, J=8.4 Hz, 1H), 4.86-4.78 (m, 1H),4.11 (d, J=6.2 Hz, 2H), 3.50-3.39 (m, 3H), 3.11-2.96 (m, 3H), 2.23 (brd, J=12.3 Hz, 2H), 2.13-1.98 (m, 5H), 1.70 (dq, J=2.8, 12.8 Hz, 2H),1.60-1.34 (m, 4H), 1.22 (br d, J=6.2 Hz, 6H), 1.11 (s, 9H). ESI[M/2+H]=318.6

Example 34. Synthesis of Isopropyl((1R,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(((((R)-1-methylpyrrolidin-2-yl)methoxy)carbonyl)amino)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.41 (d, J=2.2 Hz, 1H), 7.80-7.73 (m,2H), 7.45 (d, J=8.4 Hz, 1H), 4.86-4.81 (m, 1H), 4.67 (dd, J=3.2, 12.8Hz, 1H), 4.35 (dd, J=7.1, 12.8 Hz, 1H), 3.85-3.71 (m, 2H), 3.47 (tt,J=3.9, 11.6 Hz, 1H), 3.26 (td, J=8.1, 11.5 Hz, 1H), 3.10 (s, 3H),3.08-2.98 (m, 1H), 2.46-2.35 (m, 1H), 2.30-2.18 (m, 3H), 2.15-1.95 (m,4H), 1.72 (dq, J=3.1, 12.9 Hz, 2H), 1.43 (dq, J=3.3, 12.6 Hz, 2H), 1.25(br d, J=6.2 Hz, 6H), 1.13 (s, 9H). ESI [M+H]=636.3

Example 35. Synthesis of Isopropyl((1S,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(((((S)-1-methylpyrrolidin-2-yl)methoxy)carbonyl)amino)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.41 (d, J=2.0 Hz, 1H), 7.80-7.71 (m,2H), 7.44 (d, J=8.3 Hz, 1H), 4.86-4.79 (m, 1H), 4.67 (dd, J=3.2, 13.0Hz, 1H), 4.40-4.31 (m, 1H), 3.87-3.66 (m, 2H), 3.47 (tt, J=3.8, 11.6 Hz,1H), 3.26 (td, J=8.3, 11.4 Hz, 1H), 3.10 (s, 3H), 3.04 (tt, J=3.4, 12.0Hz, 1H), 2.48-2.33 (m, 1H), 2.30-2.16 (m, 3H), 2.15-1.92 (m, 4H), 1.72(dq, J=2.7, 12.8 Hz, 2H), 1.51-1.36 (m, 2H), 1.25 (br d, J=6.4 Hz, 6H),1.13 (s, 9H). ESI [M/2+H]=318.6

Example 36. Synthesis of Isopropyl((1R,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(((((R)-pyrrolidin-2-yl)methoxy)carbonyl)amino)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.38 (br d, J=2.2 Hz, 1H), 7.75-7.66 (m,2H), 7.41 (d, J=8.4 Hz, 1H), 4.81 (br s, 1H), 4.48 (br dd, J=3.4, 12.5Hz, 1H), 4.33 (dd, J=7.8, 12.5 Hz, 1H), 3.93 (dq, J=3.6, 8.0 Hz, 1H),3.49-3.33 (m, 3H), 3.00 (ddd, J=3.5, 8.5, 12.0 Hz, 1H), 2.30-2.18 (m,3H), 2.10-2.00 (m, 3H), 1.85 (qd, J=8.5, 12.9 Hz, 2H), 1.74-1.61 (m,2H), 1.46-1.34 (m, 2H), 1.22 (br d, J=6.2 Hz, 6H), 1.09 (s, 9H). ESI[M/2+H]=311.6

Example 37. Synthesis of Isopropyl((1S,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(((((S)-pyrrolidin-2-yl)methoxy)carbonyl)amino)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.38 (d, J=1.8 Hz, 1H), 7.79-7.68 (m,2H), 7.41 (d, J=8.2 Hz, 1H), 4.85-4.78 (m, 1H), 4.49 (dd, J=3.4, 12.5Hz, 1H), 4.33 (dd, J=7.9, 12.3 Hz, 1H), 3.94 (dq, J=3.4, 8.0 Hz, 1H),3.49-3.40 (m, 1H), 3.40-3.33 (m, 2H), 3.08-2.97 (m, 1H), 2.31-2.19 (m,3H), 2.17-2.00 (m, 4H), 1.85 (qd, J=8.5, 13.0 Hz, 1H), 1.75-1.62 (m,2H), 1.46-1.35 (m, 2H), 1.22 (br d, J=6.2 Hz, 6H), 1.10 (s, 9H). ESI[M/2+H]=311.6

Example 38. Synthesis of Isopropyl((1r,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(((oxetan-3-yloxy)carbonyl)amino)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (METHANOL-d4, 400 MHz): δ=8.33 (s, 1H), 7.63-7.72 (m, 2H), 7.38(d, J=8.4 Hz, 1H), 5.50 (br t, J=5.5 Hz, 1H), 4.92 (t, J=6.9 Hz, 2H),4.81-4.83 (m, 1H), 4.64-4.73 (m, 2H), 3.44 (br t, J=11.7 Hz, 1H), 2.99(br t, J=11.8 Hz, 1H), 2.21 (br d, J=12.3 Hz, 2H), 2.06 (br d, J=11.7Hz, 2H), 1.59-1.75 (m, 2H), 1.30-1.47 (m, 2H), 1.21 (br d, J=6.0 Hz,6H), 1.10 ppm (s, 9H). ESI [M+H]=595.1

Example 39. Synthesis of benzyl(3-(N-(tert-butyl)sulfamoyl)-4-(2-((1r,4r)-4-((isopropoxycarbonyl)amino)cyclohexyl)thiazol-5-yl)phenyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.36 (s, 1H), 7.75-7.67 (m, 2H),7.46-7.28 (m, 6H), 5.28-5.14 (m, 2H), 4.85-4.76 (m, 1H), 3.47 (br d,J=11.8 Hz, 1H), 3.01 (br s, 1H), 2.23 (br d, J=12.7 Hz, 2H), 2.07 (br d,J=14.5 Hz, 2H), 1.70 (br d, J=11.0 Hz, 2H), 1.40 (br d, J=12.7 Hz, 2H),1.22 (br d, J=6.1 Hz, 6H), 1.12 (s, 9H). ESI [M+H]=629.2

Example 40. Synthesis of 2-fluorobenzyl(3-(N-(tert-butyl)sulfamoyl)-4-(2-((1r,4r)-4-((isopropoxycarbonyl)amino)cyclohexyl)thiazol-5-yl)phenyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

1H NMR (400 MHz, METHANOL-d4) δ=8.36 (s, 1H), 7.77-7.63 (m, 2H), 7.52(t, J=7.0 Hz, 1H), 7.44-7.31 (m, 2H), 7.23-7.08 (m, 2H), 5.29 (s, 2H),4.83 (br s, 1H), 3.46 (br d, J=11.8 Hz, 1H), 3.00 (br t, J=11.8 Hz, 1H),2.22 (br d, J=12.7 Hz, 2H), 2.07 (br d, J=11.4 Hz, 2H), 1.76-1.62 (m,2H), 1.47-1.35 (m, 2H), 1.22 (br d, J=6.1 Hz, 6H), 1.12 (s, 9H). ESI[M+H]=647.2

Example 41. Synthesis of (S)-1-phenylethyl(3-(N-(tert-butyl)sulfamoyl)-4-(2-((1r,4S)-4-((isopropoxycarbonyl)amino)cyclohexyl)thiazol-5-yl)phenyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.35 (br s, 1H), 7.81-7.65 (m, 2H),7.48-7.36 (m, 5H), 7.31 (br d, J=6.8 Hz, 1H), 5.89 (br d, J=6.2 Hz, 1H),4.85 (br d, J=5.5 Hz, 1H), 3.47 (br s, 1H), 3.03 (br s, 1H), 2.24 (br d,J=11.2 Hz, 2H), 2.08 (br d, J=11.0 Hz, 2H), 1.71 (q, J=11.9 Hz, 2H),1.61 (br d, J=6.4 Hz, 3H), 1.49-1.37 (m, 2H), 1.24 (br d, J=5.4 Hz, 6H),1.13 (s, 9H). ESI [M+H]=643.2

Example 42. Synthesis of pyridin-2-ylmethyl(3-(N-(tert-butyl)sulfamoyl)-4-(2-((1r,4r)-4-((isopropoxycarbonyl)amino)cyclohexyl)thiazol-5-yl)phenyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, METHANOL-d4) δ=8.56 (br d, J=4.5 Hz, 1H), 8.38 (d,J=1.8 Hz, 1H), 7.94-7.87 (m, 1H), 7.78-7.72 (m, 2H), 7.59 (br d, J=7.8Hz, 1H), 7.44-7.38 (m, 2H), 5.32 (s, 2H), 4.85 (td, J=5.9, 12.0 Hz, 1H),3.47 (br t, J=11.8 Hz, 1H), 3.07-2.96 (m, 1H), 2.24 (br d, J=12.3 Hz,2H), 2.13-2.04 (m, 2H), 1.77-1.65 (m, 2H), 1.48-1.37 (m, 2H), 1.24 (brd, J=6.1 Hz, 6H), 1.14 (s, 9H). ESI [M+H]=630.2

Example 43. Synthesis of (R)-1-phenylethyl(3-(N-(tert-butyl)sulfamoyl)-4-(2-((1r,4R)-4-((isopropoxycarbonyl)amino)cyclohexyl)thiazol-5-yl)phenyl)carbamate

The following compound was synthesized via same method by the keyintermediate 118.

¹H NMR (400 MHz, DMSO-d6) δ=8.28 (d, J=2.2 Hz, 1H), 7.70-7.57 (m, 2H),7.47-7.24 (m, 7H), 7.05-6.84 (m, 2H), 5.82 (q, J=6.6 Hz, 1H), 4.79-4.65(m, 1H), 3.35-3.24 (m, 1H), 2.88 (tt, J=3.4, 11.9 Hz, 1H), 2.11 (br d,J=11.7 Hz, 2H), 1.89 (br d, J=9.9 Hz, 2H), 1.54 (d, J=6.6 Hz, 5H), 1.30(br s, 2H), 1.14 (d, J=6.2 Hz, 6H), 1.08-0.99 (m, 9H). ESI [M+H]=643.3

Example 44. Synthesis of trans-isopropylN-[4-[2-[4-(tert-butoxycarbonylamino)cyclohexyl]thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]carbamate

General method B, trans-isopropylN-[4-[2-[4-(tert-butoxycarbonylamino)cyclohexyl]thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]carbamate.¹H NMR (400 MHz, DMSO-d6) δ=10.06 (s, 1H), 8.32 (d, J=2.1 Hz, 1H),7.70-7.60 (m, 2H), 7.38 (d, J=8.4 Hz, 1H), 6.96 (s, 1H), 6.81 (br d,J=7.9 Hz, 1H), 4.99-4.87 (m, 1H), 3.27 (br s, 1H), 2.94-2.84 (m, 1H),2.13 (br d, J=11.7 Hz, 2H), 1.90 (br d, J=11.1 Hz, 2H), 1.63-1.49 (m,2H), 1.39 (s, 9H), 1.33 (br d, J=14.4 Hz, 2H), 1.28 (d, J=6.4 Hz, 6H),1.07 (s, 9H). ESI [M+H]=595.1

Example 45. Synthesis of tert-butyl((1r,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(pyridin-2-ylmethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 82.

¹H NMR (400 MHz, METHANOL-d4) δ=8.17 (d, J=2.2 Hz, 1H), 7.75-7.59 (m,2H), 7.43-7.22 (m, 3H), 7.21-7.03 (m, 2H), 4.53 (m, 2H), 3.43 (br t,J=11.8 Hz, 1H), 2.99 (br t, J=12.1 Hz, 1H), 2.22 (br d, J=12.7 Hz, 2H),2.06 (br d, J=10.5 Hz, 2H), 1.77-1.60 (m, 2H), 1.58-1.34 (m, 11H), 1.09(s, 9H). ESI [M+H]=643.3

Example 46. Synthesis of IsopropylN-[3-(tert-butylsulfamoyl)-4-[2-[3-(isopropoxycarbonylamino)azetidin-1-yl]thiazol-5-yl]phenyl]carbamate

Preparation of Compound 129.

A mixture of tert-butyl N-(azetidin-3-yl)carbamate; hydrochloride (72.40mg, 346.94 umol, 1.50 eq.),N-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl)phenyl] acetamide (100mg, 231.29 umol, 1 eq.), t-BuONa (66.68 mg, 693.87 umol, 3 eq.),[2-(2-aminoethyl)phenyl]-chloro-palladium;ditert-butyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane (15.88 mg,23.13 umol, 0.1 eq.) in tert-amyl alcohol (2 mL) was degassed and purgedwith N₂ for 3 times, and then the mixture was stirred at 100° C. for 12hrs under N₂ atmosphere and then concentrated. The residue was dilutedwith Ethyl acetate (20 mL) and washed with H₂O (20 mL). The organiclayer was dried and concentrated and the residue was purified byprep-TLC (EtOAc) to give tert-butylN-[1-[5-[4-acetamido-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]azetidin-3-yl]carbamate (46 mg, crude) as a yellow solid. ESI[M+H]=524.3

Preparation of Compound 130.

General method F,5-amino-2-[2-(3-aminoazetidin-1-yl)thiazol-5-yl]-N-tert-butyl-benzenesulfonamide.ESI [M+H]=382.0

Preparation of Example 46

General method D, isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[3-(isopropoxycarbonylamino)azetidin-1-yl]thiazol-5-yl]phenyl]carbamate. ¹H NMR (400 MHz,METHANOL-d4) δ=8.35 (d, J=2.0 Hz, 1H), 7.67 (dd, J=2.2, 8.4 Hz, 1H),7.45-7.32 (m, 2H), 5.04-4.94 (m, 1H), 4.92-4.88 (m, 1H), 4.73-4.63 (m,1H), 4.56 (br t, J=8.5 Hz, 2H), 4.27 (br s, 2H), 1.31 (d, J=6.2 Hz, 6H),1.27-1.19 (m, 15H). ESI [M+H]=554.2

Example 47. Synthesis of trans-oxetan-3-ylN-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate

Preparation of Compound 131.

General method F, trans-4-(5-bromothiazol-2-yl)cyclohexanamine. ESI[M+H]=260.9/262.9

Preparation of Compound 132.

To a solution of oxetan-3-ol (1.15 g, 15.51 mmol, 3 eq.) in DCE (10 mL)were added DIEA (3.34 g, 25.84 mmol, 4.50 mL, 5 eq.) and TRIPHOSGENE(1.53 g, 5.17 mmol, 1 eq.). The mixture was stirred at 25-50° C. for 1hr and then added a solution oftrans-4-(5-bromothiazol-2-yl)cyclohexanamine (1.35 g, 5.17 mmol, 1 eq.),DIEA (3.34 g, 25.84 mmol, 4.50 mL, 5 eq.) in DCE (10 mL). The mixturewas stirred at 25° C. for 0.5 hr. The mixture was concentrated and theresidue was purified by column chromatography (SiO₂, Petroleumether/Ethyl acetate=30/1 to 5:1) to give trans-oxetan-3-ylN-[4-(5-bromothiazol-2-yl)cyclohexyl]carbamate (1.5 g, crude) as a whitesolid. ESI [M+H]=363.1/361.1

Preparation of Ex. 47

General method K, trans-oxetan-3-ylN-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate.¹H NMR (400 MHz, METHANOL-d4) δ=8.23 (d, J=2.2 Hz, 1H), 7.71-7.66 (m,2H), 7.37-7.29 (m, 5H), 7.24 (br s, 1H), 5.40-5.27 (m, 2H), 4.66-4.55(m, 3H), 4.40 (s, 2H), 3.45 (br d, J=11.0 Hz, 1H), 3.00 (br t, J=11.8Hz, 1H), 2.22 (br d, J=13.0 Hz, 2H), 2.07 (br d, J=11.2 Hz, 2H),1.74-1.63 (m, 2H), 1.47-1.37 (m, 2H), 1.10 (s, 9H). ESI [M+H]=642.3

Example 48. Synthesis of oxetan-3-yl((1r,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-((isopropoxycarbonyl)amino)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 132.

¹H NMR (400 MHz, METHANOL-d4) δ=8.33 (d, J=2.0 Hz, 1H), 7.71-7.64 (m,2H), 7.36 (d, J=8.4 Hz, 1H), 5.41-5.27 (m, 2H), 4.98 (td, J=6.4, 12.6Hz, 1H), 4.87 (br s, 1H), 4.63-4.56 (m, 2H), 3.45 (br d, J=12.3 Hz, 1H),3.00 (br t, J=12.0 Hz, 1H), 2.22 (br d, J=13.0 Hz, 2H), 2.11-2.02 (m,2H), 1.74-1.63 (m, 2H), 1.42 (q, J=12.7 Hz, 2H), 1.31 (d, J=6.2 Hz, 6H),1.11 (s, 9H). ESI [M+H]=595.3

Example 49. Synthesis of trans-oxetan-3-ylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(oxetan-3-yloxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate

Preparation of Compound 133.

General method K, trans-oxetan-3-ylN-[4-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate. ESI [M+H]=509.0

Preparation of Compound 134.

General method D, trans-oxetan-3-ylN-[4-[5-[2-(tert-butylsulfamoyl)-4-[(4-nitrophenoxy)carbonylamino]phenyl]thiazol-2-yl]cyclohexyl]carbamate.ESI [M+H]=674.2

Preparation of Example 49.

General method H, trans-oxetan-3-ylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(oxetan-3-yloxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate.¹H NMR (400 MHz, DMSO-d6) δ=10.39 (s, 1H), 8.28 (d, J=2.2 Hz, 1H),7.71-7.58 (m, 2H), 7.48-7.34 (m, 2H), 6.97 (s, 1H), 5.50-5.39 (m, 1H),5.32-5.20 (m, 1H), 4.87-4.68 (m, 4H), 4.61-4.36 (m, 4H), 3.30-3.23 (m,1H), 3.01-2.82 (m, 1H), 2.12 (br d, J=11.7 Hz, 2H), 1.90 (br d, J=10.4Hz, 2H), 1.66-1.47 (m, 2H), 1.43-1.26 (m, 2H), 1.04 (s, 9H). ESI[M+H]=609.2

Example 50. Synthesis of oxetan-3-yl((1r,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(pyridin-2-ylmethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 134.

¹H NMR (400 MHz, METHANOL-d4) δ=8.49 (d, J=4.8 Hz, 1H), 8.25 (d, J=2.2Hz, 1H), 7.82 (dt, J=1.8, 7.7 Hz, 1H), 7.73-7.63 (m, 2H), 7.45 (d, J=7.9Hz, 1H), 7.38-7.26 (m, 2H), 5.39-5.28 (m, 1H), 4.84 (br s, 1H),4.70-4.56 (m, 3H), 4.53 (s, 2H), 3.44 (br t, J=12.1 Hz, 1H), 3.06-2.92(m, 1H), 2.22 (br d, J=13.2 Hz, 2H), 2.07 (br d, J=11.0 Hz, 2H),1.79-1.60 (m, 2H), 1.50-1.35 (m, 2H), 1.10 (s, 9H). ESI [M+H]=643.2

Example 51. Synthesis of oxetan-3-yl((1S,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((S)-1-phenylethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 134.

¹H NMR (400 MHz, METHANOL-d4) δ=8.19 (d, J=2.2 Hz, 1H), 7.70-7.63 (m,2H), 7.39-7.30 (m, 5H), 7.27-7.20 (m, 1H), 5.35 (t, J=5.7 Hz, 1H), 4.92(q, J=6.8 Hz, 2H), 4.84 (br s, 1H), 4.60 (t, J=6.4 Hz, 2H), 3.49-3.38(m, 1H), 3.05-2.95 (m, 1H), 2.22 (br d, J=12.3 Hz, 2H), 2.10-2.02 (m,2H), 1.74-1.61 (m, 2H), 1.49 (d, J=7.1 Hz, 3H), 1.45-1.34 (m, 2H), 1.09(s, 9H). ESI [M+H]=656.3

Example 52. Synthesis of oxetan-3-yl((1R,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((R)-1-phenylethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 134.

¹HNMR (400 MHz, METHANOL-d4) δ=8.19 (d, J=2.2 Hz, 1H), 7.75-7.61 (m,2H), 7.42-7.18 (m, 6H), 5.35 (quin, J=5.7 Hz, 1H), 5.01-4.87 (m, 3H),4.70-4.52 (m, 2H), 3.51-3.38 (m, 1H), 3.06-2.92 (m, 1H), 2.22 (br d,J=11.9 Hz, 2H), 2.07 (br d, J=11.7 Hz, 2H), 1.77-1.62 (m, 2H), 1.54-1.35(m, 5H), 1.10 (s, 9H). ESI [M+H]=656.2

Example 53. Synthesis of oxetan-3-yl((1r,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(2-fluorobenzyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 134.

¹HNMR (400 MHz, METHANOL-d4) δ=8.23 (d, J=2.4 Hz, 1H), 7.75-7.63 (m,2H), 7.46-7.24 (m, 3H), 7.20-7.03 (m, 2H), 5.43-5.28 (m, 1H), 4.85 (brs, 2H), 4.67-4.56 (m, 2H), 4.47 (s, 2H), 3.53-3.37 (m, 1H), 3.08-2.93(m, 1H), 2.29-1.99 (m, 4H), 1.78-1.61 (m, 2H), 1.53-1.35 (m, 2H), 1.11(s, 9H). ESI [M+H]=660.2

Example 54. Synthesis of oxetan-3-yl((1R,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((R)-1-(2-fluorophenyl)ethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 134.

¹H NMR (400 MHz, METHANOL-d4) δ=8.20 (d, J=2.3 Hz, 1H), 7.73-7.65 (m,2H), 7.45-7.39 (m, 1H), 7.38-7.26 (m, 2H), 7.21-7.04 (m, 2H), 5.37(quin, J=5.7 Hz, 1H), 5.20 (q, J=7.0 Hz, 1H), 4.89-4.86 (m, 2H),4.65-4.59 (m, 2H), 3.52-3.41 (m, 1H), 3.08-2.96 (m, 1H), 2.24 (br d,J=12.2 Hz, 2H), 2.13-2.05 (m, 2H), 1.76-1.64 (m, 2H), 1.52 (d, J=7.0 Hz,3H), 1.47-1.39 (m, 2H), 1.12 (s, 9H). ESI [M+H]=674.2

Example 55. Synthesis of oxetan-3-yl((1S,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-((S)-1-(2-fluorophenyl)ethyl)ureido)phenyl)thiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 134.

¹HNMR (400 MHz, METHANOL-d4) δ=8.17 (d, J=2.2 Hz, 1H), 7.75-7.59 (m,2H), 7.43-7.22 (m, 3H), 7.21-7.03 (m, 2H), 5.39-5.28 (m, 1H), 5.17 (q,J=7.0 Hz, 1H), 4.90-4.87 (m, 1H), 4.84 (s, 1H), 4.66-4.54 (m, 2H), 3.43(br t, J=11.8 Hz, 1H), 2.99 (br t, J=12.1 Hz, 1H), 2.22 (br d, J=12.7Hz, 2H), 2.06 (br d, J=10.5 Hz, 2H), 1.77-1.60 (m, 2H), 1.58-1.34 (m,5H), 1.09 (s, 9H). ESI [M+H]=674.2

Example 56. Synthesis of 4-piperidylmethylN-[3-(tert-butylsulfamoyl)-4-[2-[5-(isopropoxycarbonylamino)-3-methoxy-2-pyridyl]thiazol-5-yl]phenyl]carbamate

Preparation of Compound 136.

Na (8.75 g, 380.60 mmol, 9.02 mL, 1.51 eq.) was added into MeOH (300 mL)portionwise and after Na was dissolved, the mixture was concentrated todryness. The resulting gray solid (NaOMe) was added into DMF (300 mL)and 3-chloropyridine-2-carbonitrile (35 g, 252.61 mmol, 1 eq.) was addedat 0° C. The reaction mixture was stirred at 20° C. for 12 hrs and thendiluted with H₂O (800 mL) and filtered. The cake dried to give3-methoxypyridine-2-carbonitrile (25 g, crude) as a white solid. ESI[M+H]=135.1

Preparation of Compound 137

To a solution of 3-methoxypyridine-2-carbonitrile (24.5 g, 182.65 mmol,1 eq.) in DCM (450 mL) was added a mixture of Bu₄NNO3 (83.30 g, 273.59mmol, 1.5 eq.) and TFAA (57.54 g, 273.98 mmol, 38.11 mL, 1.5 eq.) in DCM(150 mL) dropwise at 0° C. The mixture was stirred at 20° C. for 12 hrsand then poured into sat.aq.NaHCO₃ (300 mL) at 0° C. and the organiclayer was dried over Na₂SO₄, filtered and concentrated. The residue waspurified by column chromatography (SiO₂, Petroleum ether/Ethylacetate=20/1 to 10:1) to give 3-methoxy-5-nitro-pyridine-2-carbonitrile(27 g, crude) as a white solid. ¹H NMR (400 MHz, CHLOROFORM-d) δ=9.06(d, J=2.2 Hz, 1H), 8.13 (d, J=2.2 Hz, 1H), 4.16-4.08 (m, 3H). ESI[M+H]=180.1

General Method G for Preparation of Compound 138.

To a solution of 3-methoxy-5-nitro-pyridine-2-carbonitrile (27 g, 151mmol, 1 eq.) in THF (100 mL)/EtOH (500 mL) were added Fe (4 g, 754 mmol,5 eq.) and a solution of NH₄Cl (24.2 g, 452 mmol, 3 eq.) in H₂O (50 mL).The mixture was stirred at 80° C. for 30 mins, then diluted with THF(500 mL) and filtered. The filtrate was concentrated, diluted with H₂O(500 mL) and then extracted with DCM (400 mL×3). The combined organiclayers were dried over Na₂SO₄, filtered and concentrated to give5-amino-3-methoxy-pyridine-2-carbonitrile (14.5 g, crude) as a paleyellow solid. ESI [M+H]=150.1

Preparation of Compound 139.

General method D, isopropyl N-(6-cyano-5-methoxy-3-pyridyl)carbamate. ¹HNMR (400 MHz, METHANOL-d4) δ=8.13 (d, J=2.2 Hz, 1H), 7.95 (d, J=1.8 Hz,1H), 5.04-4.95 (m, 1H), 3.96 (s, 3H), 1.31 (d, J=6.2 Hz, 6H). ESI[M+H]=236.1

Preparation of Compound 140.

To a solution of isopropyl N-(6-cyano-5-methoxy-3-pyridyl)carbamate (16g, 68.02 mmol, 1 eq.) in DMF (200 mL) were added NaHS (19.06 g, 340.08mmol, 5 eq.) and then MgCl₂ (19.43 g, 204.05 mmol, 8.37 mL, 3 eq.). Themixture was stirred at 25° C. for 12 hrs and then poured into H₂O (500ml) and extracted with DCM (40 mL*3). The combined organic layers weredried over Na₂SO₄, filtered and concentrated. The residue was washedwith a solution (Petroleum ether:EtOAc=8:1), filtered and the filtercake was dried to give isopropylN-(6-carbamothioyl-5-methoxy-3-pyridyl)carbamate (21 g, crude) as ayellow solid. ¹H NMR (400 MHz, DMSO-d6) δ=9.92-9.84 (m, 2H), 9.38 (br s,1H), 8.08 (d, J=1.8 Hz, 1H), 7.66 (s, 1H), 4.89 (spt, J=6.3 Hz, 1H),3.74 (s, 3H), 1.24 (d, J=6.1 Hz, 6H). ESI [M+H]=270.0

Preparation of Compound 141.

General method M, isopropylN-(5-methoxy-6-thiazol-2-yl-3-pyridyl)carbamate ¹H NMR (400 MHz,DMSO-d6) δ=10.07 (s, 1H), 8.26 (d, J=2.0 Hz, 1H), 7.88 (d, J=3.1 Hz,1H), 7.85 (d, J=1.3 Hz, 1H), 7.71 (d, J=3.3 Hz, 1H), 4.91 (spt, J=6.2Hz, 1H), 3.88 (s, 3H), 1.26 (d, J=6.4 Hz, 6H). ESI [M+H]=294.1

Preparation of Compound 142.

General method J, isopropylN-[6-(5-bromothiazol-2-yl)-5-methoxy-3-pyridyl] carbamate. ¹H NMR (400MHz, DMSO-d6) δ=10.17 (s, 1H), 8.28 (d, J=1.8 Hz, 1H), 8.08-7.86 (m,2H), 4.99 (br s, 1H), 3.93 (s, 3H), 1.29 (d, J=6.4 Hz, 6H). ESI[M+H]=371.8/373.8

Preparation of Compound 143.

General method B, isopropylN-[6-[5-[4-acetamido-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-5-methoxy-3-pyridyl]carbamate. ESI [M+H]=562.0

Preparation of Compound 144.

General method F, isopropylN-[6-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-5-methoxy-3-pyridyl]carbamate. ESI [M+H]=520.2

Preparation of Compound 145.

General method D,(4-nitrophenyl)N-[3-(tert-butylsulfamoyl)-4-[2-[5-(isopropoxycarbonylamino)-3-methoxy-2-pyridyl]thiazol-5-yl]phenyl]carbamate.ESI [M+H]=685.1

Preparation of Compound 146.

General method H, tert-butyl4-[[3-(tert-butylsulfamoyl)-4-[2-[5-(isopropoxycarbonylamino)-3-methoxy-2-pyridyl]thiazol-5-yl]phenyl]carbamoyloxymethyl]piperidine-1-carboxylate.ESI [M+H]=761.4

Preparation of Ex. 56

General method C, 4-piperidylmethylN-[3-(tert-butylsulfamoyl)-4-[2-[5-(isopropoxycarbonylamino)-3-methoxy-2-pyridyl]thiazol-5-yl]phenyl]carbamate.¹H NMR (400 MHz, DMSO-d6) δ=10.19-10.10 (m, 2H), 8.35 (d, J=2.1 Hz, 1H),8.30 (d, J=1.8 Hz, 1H), 7.89 (s, 2H), 7.68 (dd, J=2.0, 8.4 Hz, 1H), 7.46(d, J=8.4 Hz, 1H), 7.12 (br s, 1H), 4.95 (td, J=6.2, 12.5 Hz, 1H), 3.98(d, J=6.5 Hz, 2H), 3.93 (s, 3H), 3.00 (br d, J=12.1 Hz, 2H), 2.70-2.56(m, 2H), 1.77 (br d, J=4.0 Hz, 1H), 1.67 (br d, J=12.1 Hz, 2H), 1.30 (d,J=6.2 Hz, 6H), 1.16 (br dd, J=3.4, 12.0 Hz, 2H), 1.09 (s, 9H). ESI[M+H]=661.3

Example 57 Synthesis of trans-isopropylN-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]-4-fluoro-thiazol-2-yl]cyclohexyl]carbamate

Preparation of Compound 147

To a solution of trans-isopropylN-[4-(5-bromothiazol-2-yl)cyclohexyl]carbamate (1.50 g, 4.32 mmol, 1eq.) in ACN (20 mL) was added1-(chloromethyl)-4-fluoro-1,4-diazoniabicyclo[2.2.2]octane;ditetrafluoroborate (3.06 g, 8.64 mmol, 2 eq.) and the mixture wasstirred at 80° C. for 12 hrs. The mixture was then concentrated and theresidue was purified by column chromatography (SiO₂, Petroleumether/Ethyl acetate=3/1 to 3:1) to give trans-isopropylN-[4-(5-bromo-4-fluoro-thiazol-2-yl) cyclohexyl]carbamate (0.2 g, 547.55umol, 12.68% yield) as yellow gum. ESI [M+H]=365.1/367.1

Preparation of Ex. 57

General method B, trans-isopropylN-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]-4-fluoro-thiazol-2-yl]cyclohexyl]carbamate.¹H NMR (400 MHz, METHANOL-d4) δ=8.24 (d, J=2.0 Hz, 1H), 7.67 (dd, J=2.0,8.4 Hz, 1H), 7.37-7.29 (m, 5H), 7.24 (br d, J=2.6 Hz, 1H), 4.82-4.77 (m,1H), 4.40 (s, 2H), 3.42 (br t, J=11.6 Hz, 1H), 2.87 (br t, J=12.0 Hz,1H), 2.20 (br d, J=12.3 Hz, 2H), 2.05 (br d, J=11.0 Hz, 2H), 1.69-1.57(m, 2H), 1.43-1.30 (m, 2H), 1.21 (br d, J=5.7 Hz, 6H), 1.15 (s, 9H). ESI[M+H]=646.2

Example 58 Synthesis of isopropyl((1r,4r)-4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-((isopropoxycarbonyl)amino)phenyl)-4-fluorothiazol-2-yl)cyclohexyl)carbamate

The following compound was synthesized via same method by the keyintermediate 147.

¹H NMR (400 MHz, METHANOL-d4) δ=8.35 (s, 1H), 7.69 (br d, J=8.1 Hz, 1H),7.37 (d, J=8.3 Hz, 1H), 5.01 (td, J=6.1, 12.4 Hz, 1H), 4.83 (br s, 1H),3.53-3.39 (m, 1H), 2.96-2.85 (m, 1H), 2.23 (br d, J=12.3 Hz, 2H), 2.08(br d, J=10.5 Hz, 2H), 1.75-1.60 (m, 2H), 1.47-1.38 (m, 2H), 1.34 (d,J=6.2 Hz, 6H), 1.24 (br d, J=6.0 Hz, 6H), 1.18 (s, 9H). ESI [M+H]=599.2

Example 59 Synthesis of trans-isopropylN-[6-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]tetrahydropyran-3-yl]carbamate

Preparation of Compound 149

To a solution of ethyl 2-oxoacetate (250 g, 1.22 mol, 1 eq.) in Tol.(1.5 L), were added buta-1,3-diene (92.72 g, 1.71 mol, 149.55 mL, 1.4eq.) and 2,6-ditert-butyl-4-methyl-phenol (5.40 g, 24.49 mmol, 0.02eq.). The mixture was stirred at 170° C. for 8 hrs in high pressure tubeunder 1 MPa and then concentrated. The residue was purified by silicagel chromatography (Petroleum ether/Ethyl acetate=100:1-10:1) to affordethyl 3,6-dihydro-2H-pyran-2-carboxylate (26 g, 166.48 mmol, 13.60%yield) as yellow oil. ¹H NMR (400 MHz, CHLOROFORM-d) δ=5.82-5.74 (m,1H), 5.71-5.64 (m, 1H), 4.36-4.27 (m, 1H), 4.23-4.12 (m, 4H), 2.39-2.23(m, 2H), 1.24 (t, J=7.1 Hz, 3H).

Preparation of Compound 150

General method O, 3,6-dihydro-2H-pyran-2-carboxylic acid. ¹H NMR (400MHz, CHLOROFORM-d) δ=5.88 (qdd, J=2.3, 5.1, 10.2 Hz, 1H), 5.80-5.71 (m,1H), 4.43-4.19 (m, 3H), 2.53-2.31 (m, 2H)

Preparation of Compound 151

General method N, 3,6-dihydro-2H-pyran-2-carboxamide. ¹H NMR (400 MHz,CHLOROFORM-d) δ=6.50 (br s, 1H), 5.89-5.76 (m, 1H), 5.67 (br d, J=9.8Hz, 1H), 5.50 (br s, 1H), 4.22 (br s, 2H), 3.97 (dd, J=3.9, 10.8 Hz,1H), 2.49-2.35 (m, 1H), 2.28-2.09 (m, 1H)

Preparation of Compound 152

General method L, 3,6-dihydro-2H-pyran-2-carbothioamide. ESI [M+H]=144.1

Preparation of Compound 153

General method M, 2-(3,6-dihydro-2H-pyran-2-yl)thiazole. ¹H NMR (400MHz, CHLOROFORM-d) δ=7.69 (d, J=2.9 Hz, 1H), 7.26 (d, J=2.9 Hz, 1H),5.93-5.83 (m, 1H), 5.70-5.63 (m, 1H), 4.86 (dd, J=3.9, 9.8 Hz, 1H), 4.33(br s, 2H), 2.61-2.37 (m, 2H). ESI [M+H]=168.1

Preparation of Compound 154

To a solution of 2-(3,6-dihydro-2H-pyran-2-yl)thiazole (13 g, 77.74mmol, 1 eq.) in THF (150 mL), was added BH₃-Me₂S (10 M, 15.55 mL, 2 eq.)at 0° C. dropwise and the mixture was stirred at 26° C. for 2 hrs. Thenthe mixture was quenched by NaOH (62.19 g, 1.55 mol, 20 eq.) in H₂O (150mL) slowly at 0° C. followed by addition of H₂O₂(264.42 g, 2.33 mol,224.09 mL, 30% purity, 30 eq.). The mixture was stirred at 26° C. foranother 12 hrs. The mixture was quenched by sat.aq.Na₂SO₃ solution (1 L)and extracted with EtOAc (1 L*2). The combined organic phased was driedover Na₂SO₄, filtered and concentrated. The residue was purified bycolumn (Petroleum ether:EtOAc=5:1-1:1) to afford6-thiazol-2-yltetrahydropyran-3-ol (7 g, crude) as a yellow solid. ESI[M+H]=186.2

Preparation of Compound 155

To a solution of 6-thiazol-2-yltetrahydropyran-3-ol (7 g, 37.79 mmol, 1eq.) in DCM (100 mL), were added TEA (7.65 g, 75.58 mmol, 10.52 mL, 2eq.) and methanesulfonyl chloride (6.49 g, 56.68 mmol, 4.39 mL, 1.5 eq.)dropwise at 0° C. and the mixture was stirred at 26° C. for 2 hrs. Themixture was diluted with DCM (100 mL) and washed with water (200 mL).The organic phase was dried over Na₂SO₄, filtered and concentrated togive crude (6-thiazol-2-yltetrahydropyran-3-yl) methanesulfonate (7 g,crude) as yellow oil which can be used directly.

To a solution of (6-thiazol-2-yltetrahydropyran-3-yl) methanesulfonate(7 g, 26.58 mmol, 1 eq.) in DMF (60 mL), was added azidosodium (8.64 g,132.91 mmol, 5 eq.) and the mixture was stirred at 80° C. for 12 hrs andthen poured into sat.aq.Na₂CO₃ (500 mL) and extracted with EtOAc (200mL*3). The combined organic phase was washed with brine (200 mL) anddried over Na₂SO₄, filtered and concentrated to give crude2-(5-azidotetrahydropyran-2-yl)thiazole (5 g, crude) as yellow oil whichcan be used without any purification.

Preparation of Compound 156

To a solution of 2-(5-azidotetrahydropyran-2-yl)thiazole (5 g, 23.78mmol, 1 eq.) in THF (80 mL) and H₂O (40 mL), was added PPh₃ (9.36 g,35.67 mmol, 1.5 eq.). The mixture was stirred at 50° C. for 12 hrs andthen poured into 4N HCl solution (100 mL) and extracted with EtOAc (50mL*2). Then the aqueous phase was bacified by sat.aq.Na₂CO₃ until pH>12and extracted with a solution (DCM/MeOH=5:1) (100 mL*3). The combinedorganic phase was dried over Na₂SO₄, filtered and concentrated to givecrude 6-thiazol-2-yltetrahydropyran-3-amine (3 g, crude) as yellow oil.ESI [M+H]=185.2

Preparation of Compound 157

General method D, trans-isopropylN-(6-thiazol-2-yltetrahydropyran-3-yl)carbamate. ¹H NMR (400 MHz,METHANOL-d4) δ=7.74 (d, J=3.3 Hz, 1H), 7.55 (d, J=3.3 Hz, 1H), 4.86-4.77(m, 2H), 4.64 (dd, J=2.4, 10.8 Hz, 1H), 4.12 (ddd, J=2.0, 4.6, 10.8 Hz,1H), 3.75-3.55 (m, 1H), 2.32-2.20 (m, 1H), 2.16-2.03 (m, 1H), 1.81-1.54(m, 2H), 1.22 (br d, J=6.2 Hz, 6H). ESI [M+H]=271.2

Note: Cpd.157 was purified by prep-TLC and then prep-HPLC to separateout other isomers.

Preparation of Compound 158

General method J, trans-isopropylN-[6-(5-bromothiazol-2-yl)tetrahydropyran-3-yl]carbamate. ESI[M+H]=351.1/349.1

Preparation of Compound Ex. 59

General method B, trans-isopropylN-[6-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]tetrahydropyran-3-yl]carbamate.¹H NMR (400 MHz, METHANOL-d4) δ=8.24 (d, J=2.2 Hz, 1H), 7.74 (s, 1H),7.68 (dd, J=2.2, 8.4 Hz, 1H), 7.37-7.29 (m, 5H), 7.27-7.19 (m, 1H),4.84-4.78 (m, 2H), 4.64 (dd, J=2.0, 11.0 Hz, 1H), 4.40 (s, 2H), 4.12 (brdd, J=3.2, 10.7 Hz, 1H), 3.69-3.58 (m, 1H), 2.33-2.26 (m, 1H), 2.13 (brd, J=10.4 Hz, 1H), 1.82-1.71 (m, 1H), 1.69-1.57 (m, 1H), 1.26-1.19 (m,6H), 1.10 (s, 9H). ESI [M+H]=630.3

Preparation of Compound Ex. 59A and Ex. 59B

Ex. 59 was further separated by SFC (condition: Instrument: Thar SFC80preparative SFC; Column: Chiralpak IC-H 250*30 mm i.d. 5u; Mobile phase:A for CO2 and B for MeOH(0.1% NH₃.H₂O); Gradient: B %=42%; Flow rate 70g/min; Wavelength: 220 nm; Column temperature: 40° C.; System backpressure: 100 bar to give Ex. 59A. ¹H NMR (400 MHz, METHANOL-d4) δ=8.27(d, J=2.2 Hz, 1H), 7.77 (s, 1H), 7.72 (dd, J=2.3, 8.3 Hz, 1H), 7.40-7.33(m, 5H), 7.30-7.23 (m, 1H), 4.89-4.79 (m, 2H), 4.67 (dd, J=2.3, 11.0 Hz,1H), 4.43 (s, 2H), 4.16 (br dd, J=3.0, 10.9 Hz, 1H), 3.67 (br t, J=10.9Hz, 1H), 2.33 (br dd, J=2.6, 13.1 Hz, 1H), 2.16 (br d, J=10.6 Hz, 1H),1.88-1.58 (m, 2H), 1.31-1.22 (m, 6H), 1.13 (s, 9H). ESI [M+H]=630.2

Ex. 59B:

¹H NMR (400 MHz, METHANOL-d4) δ=8.25 (d, J=2.2 Hz, 1H), 7.76 (s, 1H),7.70 (dd, J=2.2, 8.3 Hz, 1H), 7.37 (s, 1H), 7.36-7.31 (m, 4H), 7.28-7.18(m, 1H), 4.87-4.74 (m, 1H), 4.65 (br d, J=9.2 Hz, 1H), 4.41 (s, 2H),4.19-4.04 (m, 1H), 3.63 (br d, J=10.5 Hz, 1H), 3.34 (br s, 1H), 2.31 (brd, J=13.2 Hz, 1H), 2.13 (br d, J=10.1 Hz, 1H), 1.84-1.72 (m, 1H),1.71-1.58 (m, 1H), 1.22 (d, J=6.1 Hz, 6H), 1.11 (s, 9H). ESI [M+H]=630.3

Example 60 Synthesis of trans-isopropylN-[6-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxycarbonylamino)phenyl]thiazol-2-yl]tetrahydropyran-3-yl]carbamate

Preparation of Ex. 60

General method B, trans-isopropylN-[6-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxycarbonylamino)phenyl]thiazol-2-yl]tetrahydropyran-3-yl]carbamate.¹H NMR (400 MHz, METHANOL-d4) δ=8.37 (d, J=2.0 Hz, 1H), 7.79 (s, 1H),7.70 (dd, J=2.4, 8.3 Hz, 1H), 7.40 (d, J=8.3 Hz, 1H), 5.01 (spt, J=6.3Hz, 1H), 4.87-4.79 (m, 1H), 4.68 (dd, J=2.4, 11.2 Hz, 1H), 4.16 (dd,J=2.9, 10.8 Hz, 1H), 3.72-3.60 (m, 1H), 3.38-3.34 (m, 1H), 2.39-2.29 (m,1H), 2.16 (br d, J=12.2 Hz, 1H), 1.89-1.74 (m, 1H), 1.72-1.59 (m, 1H),1.39-1.21 (m, 12H), 1.14 (s, 9H). ESI [M+H]=583.3

Preparation of Compound Ex. 60A and Ex. 60B

Ex. 60 was further separated by SFC (condition: Instrument: Thar SFC80preparative SFC; Column: Chiralpak IC-H 250*30 mm i.d. 5u; Mobile phase:A for C02 and B for MeOH(0.1% NH₃.H₂O); Gradient: B %=38%; Flow rate: 65g/min; Wavelength: 220 nm; Column temperature: 40° C.; System backpressure: 100 bar) to give Ex. 60A: ¹H NMR (400 MHz, METHANOL-d4) δ=8.37(d, J=2.2 Hz, 1H), 7.78 (s, 1H), 7.70 (dd, J=2.0, 8.4 Hz, 1H), 7.40 (d,J=8.3 Hz, 1H), 5.05-4.95 (m, 1H), 4.90-4.81 (m, 1H), 4.67 (dd, J=2.4,11.1 Hz, 1H), 4.70-4.64 (m, 1H), 4.16 (br dd, J=2.9, 10.9 Hz, 1H),3.73-3.62 (m, 1H), 2.33 (br dd, J=2.8, 13.2 Hz, 1H), 2.16 (br d, J=11.9Hz, 1H), 1.86-1.74 (m, 1H), 1.70-1.61 (m, 1H), 1.34 (d, J=6.2 Hz, 6H),1.25 (br d, J=6.1 Hz, 6H), 1.14 (s, 9H). ESI [M+H]=583.3

Ex. 60B:

¹H NMR (400 MHz, METHANOL-d4) δ=8.34 (d, J=2.2 Hz, 1H), 7.80-7.72 (m,1H), 7.67 (dd, J=2.0, 8.6 Hz, 1H), 7.36 (d, J=8.3 Hz, 1H), 4.98 (spt,J=6.2 Hz, 1H), 4.85-4.75 (m, 1H), 4.65 (dd, J=2.4, 11.2 Hz, 1H), 4.13(br dd, J=3.1, 11.0 Hz, 1H), 3.72-3.56 (m, 1H), 3.33 (s, 1H), 2.30 (brdd, J=2.6, 13.2 Hz, 1H), 2.13 (br d, J=11.4 Hz, 1H), 1.85-1.71 (m, 1H),1.69-1.57 (m, 1H), 1.31 (d, J=6.1 Hz, 6H), 1.27-1.17 (m, 6H), 1.11 (s,9H). ESI [M+H]=583.2

Example 61 Synthesis of[1-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-4-bicyclo[2.2.2]octanyl]N-isopropylcarbamate

Preparation of Compound 160.

To a solution of methyl 4-hydroxybicyclo[2.2.2]octane-1-carboxylate (0.4g, 2.17 mmol, 1 eq.) in DCM (10 mL) were added TMSCl (23.59 mg, 217.12umol, 0.1 eq.) and 2-isocyanatopropane (554.33 mg, 6.51 mmol, 3 eq.).The mixture was stirred at 25° C. for 12 hrs and then washed with 1 NHCl (20 mL) and sat.aq.Na₂CO₃ (20 mL). The organic layer was dried overNa₂SO₄, filtered and concentrated to give methyl4-(isopropylcarbamoyloxy)bicyclo[2.2.2]octane-1-carboxylate (0.45 g,crude) as a yellow gum. ¹H NMR (400 MHz, CHLOROFORM-d) δ=4.34 (br s,1H), 3.72-3.61 (m, 1H), 3.56 (s, 3H), 1.94 (br d, J=7.7 Hz, 6H),1.90-1.81 (m, 6H), 1.08-1.00 (m, 6H). ESI [M+H]=269.9

Preparation of Compound 161.

General method O,4-(isopropylcarbamoyloxy)bicyclo[2.2.2]octane-1-carboxylic acid. ¹H NMR(400 MHz, CHLOROFORM-d) δ=4.32 (br s, 1H), 3.71-3.60 (m, 1H), 1.93 (brs, 6H), 1.88 (br d, J=9.3 Hz, 6H), 1.05 (d, J=6.5 Hz, 6H). ESI[M+H]=256.0

Preparation of Compound 162.

General method N,(1-carbamoyl-4-bicyclo[2.2.2]octanyl)N-isopropylcarbamate. ¹H NMR (400MHz, CHLOROFORM-d) δ=5.56-5.31 (m, 2H), 4.34 (br s, 1H), 3.72-3.57 (m,1H), 1.96 (br s, 6H), 1.97-1.93 (m, 1H), 1.92-1.80 (m, 6H), 1.05 (d,J=6.6 Hz, 6H). ESI [M+H]=255.3

Preparation of Compound 163.

General method L,(1-carbamothioyl-4-bicyclo[2.2.2]octanyl)N-isopropylcarbamate. ESI[M+H]=271.3

Preparation of Compound 164.

General method M,(1-thiazol-2-yl-4-bicyclo[2.2.2]octanyl)N-isopropylcarbamate. ESI[M+H]=295.0

Preparation of Compound 165

General method J, [1-(5-bromothiazol-2-yl)-4-bicyclo[2.2.2]octanyl]N-isopropylcarbamate. ESI [M+H]=372.8/374.8

Preparation of Ex. 61.

General method K,[1-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-4-bicyclo[2.2.2]octanyl]N-isopropylcarbamate. ¹H NMR (400 MHz, METHANOL-d4) δ=8.26 (d, J=2.2 Hz,1H), 7.74-7.68 (m, 2H), 7.38-7.32 (m, 4H), 7.30-7.23 (m, 1H), 4.43 (s,2H), 3.71-3.61 (m, 1H), 2.18 (s, 12H), 1.16-1.09 (m, 15H). ESI[M+H]=654.3

Example 62 Synthesis of4-(5-(2-(N-(tert-butyl)sulfamoyl)-4-(3-(pyridin-2-ylmethyl)ureido)phenyl)thiazol-2-yl)bicyclo[2.2.2]octan-1-ylisopropylcarbamate

The following compound was synthesized via same method by the keyintermediate 165.

¹H NMR (400 MHz, METHANOL-d4) δ=8.77 (br s, 1H), 8.62-8.52 (m, 1H), 8.34(br s, 1H), 8.12-8.03 (m, 1H), 7.96 (br d, J=5.5 Hz, 1H), 7.79-7.66 (m,2H), 7.43-7.35 (m, 1H), 4.78 (br s, 2H), 3.67 (br dd, J=6.2, 13.0 Hz,1H), 2.19 (br s, 12H), 1.17-1.09 (m, 15H). ESI [M/2+H]=328.2

Example 63 Synthesis of trans-isopropylN-[6-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxycarbonylamino)phenyl]thiazol-2-yl]-3-piperidyl]carbamate

General Method I for Preparation of Compound 167.

To a solution of 5-hydroxypyridine-2-carboxylic acid (60 g, 431 mmol, 1eq.) in AcOH (200 mL)/H₂O (600 mL) was added wet Pd/C (2 g, 10% content)under N₂. The suspension was degassed under vacuum and purged with H₂several times. The mixture was stirred under H₂ (50 psi) at 50° C. for12 hrs, then filtered and concentrated to give5-hydroxypiperidine-2-carboxylic acid (62.61 g, crude) as a yellow oil.ESI [M+H]=146.5

Preparation of Compound 168

To a mixture of 5-hydroxypiperidine-2-carboxylic acid (62 g, 427.13mmol, 1 eq.) and Boc₂O (102.54 g, 469.84 mmol, 107.94 mL, 1.1 eq.) indioxane (500 mL) was added NaOH (34.17 g, 854.25 mmol, 2 eq.) and themixture was stirred at 25° C. for 18 hrs. The mixture was thenconcentrated to remove dioxane and the pH was adjusted to 2-3 byaddition of 1N HCl solution. The aqueous phase was extracted with2-Me-THF (500 mL*3). The combined organic layers was dried over Na₂SO₄,filtered and concentrated to give1-tert-butoxycarbonyl-5-hydroxy-piperidine-2-carboxylic acid (40 g,crude) as yellow oil. ESI [M+Na]=267.9

Preparation of Compound 169

General method A, O₂-benzyl O₁-tert-butyl5-hydroxypiperidine-1,2-dicarboxylate. ESI [M+Na+]=358.0

Preparation of Compound 170

A mixture of O₂-benzyl O1-tert-butyl5-hydroxypiperidine-1,2-dicarboxylate (10 g, 29.82 mmol, 1 eq.), TEA(3.62 g, 35.78 mmol, 4.98 mL, 1.2 eq.) in DCM (100 mL) was addedmethanesulfonyl chloride (4.10 g, 35.78 mmol, 2.77 mL, 1.2 eq.) at 0° C.and the mixture was stirred at 25° C. 1 hr. The mixture was then washedwith H₂O (50 mL) and the organic layer was dried and concentrated. Theresidue was purified by column chromatography (SiO₂, Petroleumether/Ethyl acetate=40/1 to 5:1) to afford O₂-benzyl 01-tert-butyl5-methylsulfonyloxypiperidine-1,2-dicarboxylate (9.5 g, 18.98 mmol,63.65% yield, 82.6% purity) as yellow oil.

A mixture of O₂.benzyl O1-tert-butyl5-methylsulfonyloxypiperidine-1,2-dicarboxylate (9.5 g, 22.98 mmol, 1eq.), NaN₃ (8.96 g, 137.85 mmol, 6 eq.) in DMF (20 mL) was stirred at100° C. for 4 hrs. Then the mixture was quenched with sat.aq.Na₂SO₃ (30mL) and extracted with EtOAc (100 mL*3). The combined organic layerswere washed with brine (40 mL*3), dried over Na₂SO₄, filtered andconcentrated to give O₂-benzyl O₁-tert-butyl5-azidopiperidine-1,2-dicarboxylate (8.82 g, crude) as yellow oil.

Preparation of Compound 171

A mixture of O₂-benzyl O₁-tert-butyl 5-azidopiperidine-1,2-dicarboxylate(8 g, 22.20 mmol, 1 eq.), triphenylphosphane (8.73 g, 33.30 mmol, 1.5eq.), in H₂O (50 mL) and THF (50 mL) was stirred at 45° C. for 4 hrs.The mixture was concentrated to remove the THF and pH was adjust to 2-3by addition of 1N HCl solution. The aqueous phase was extracted withMTBE (30 mL) and then aqueous phase was basified to adjust pH to 9 andextracted with EtOAc (50 mL*3). The combined organic layers were driedover Na₂SO₄ filtered and concentrated to give O2-benzyl O1-tert-butyl5-aminopiperidine-1,2-dicarboxylate (6 g, crude) as yellow oil. ESI[M+H]=335.2

Preparation of Compound 172

General method D, O₂-benzyl O₁-tert-butyl5-(isopropoxycarbonylamino)piperidine-1,2-dicarboxylate. ESI [M+H]=421.2

Preparation of Compound 173

General method O,1-tert-butoxycarbonyl-5-isopropoxycarbonyloxy-piperidine-2-carboxylicacid. ESI [M+H]=331.2

Preparation of Compound 174

A mixture of1-tert-butoxycarbonyl-5-(isopropoxycarbonylamino)piperidine-2-carboxylicacid (1.3 g, 3.93 mmol, 1 eq.), NH₄Cl (315.73 mg, 5.90 mmol, 1.5 eq.),TEA (1.19 g, 11.80 mmol, 1.64 mL, 3 eq.) and HBTU (1.64 g, 4.33 mmol,1.1 eq.) in ACN (10 mL) as stirred at 25° C. for 2 hrs and thenconcentrated. The mixture was then poured into H₂O (20 mL) and extractedwith EtOAc (20 ml*3). The combined organic layers were dried overNa₂SO₄, filtered and concentrated. The mixture was purified byprep-HPLC(column: Phenomenex Gemini C18 250*50 10u; mobile phase: [water(0.05% ammonia hydroxide v/v)-ACN]; B %: 10%-40%,20 min) to givetert-butyl2-carbamoyl-5-(isopropoxycarbonylamino)piperidine-1-carboxylate (1 g,3.01 mmol, 76.38% yield, 99% purity) as a white solid. ESI [M+H]=330.2

Preparation of Compound 175

General method L, trans-tert-butyl2-carbamothioyl-5-(isopropoxycarbonylamino) piperidine-1-carboxylate.ESI [M+H]=346.1

Preparation of Compound 176

To a solution of trans-tert-butyl2-carbamothioyl-5-(isopropoxycarbonylamino) piperidine-1-carboxylate(650 mg, 1.88 mmol, 1.0 eq.) in tol. (20 mL) were added BUFFER (784.76mg, 2.82 mmol, 1.5 eq.) and 2-chloroacetaldehyde (3.69 g, 18.82 mmol,3.03 mL, 10 eq.). The mixture was stirred at 100° C. for 1.5 hrs andthen poured into water (20 mL) and extracted with EtOAc (10 mL×3). Thecombined organic phase was dried over Na₂SO₄, filtered and concentrated.The residue was purified by basic prep-HPLC to give trans-tert-butyl5-(isopropoxycarbonylamino)-2-thiazol-2-yl-piperidine-1-carboxylate (80mg, 216.52 umol, 11.51% yield) as a yellow solid. ¹H NMR (400 MHz,METHANOL-d4) δ=7.75 (d, J=3.4 Hz, 1H), 7.58 (d, J=2.9 Hz, 1H), 5.60 (brs, 1H), 4.95-4.90 (m, 1H), 4.23 (br d, J=14.7 Hz, 1H), 3.68 (br s, 1H),3.11-2.94 (m, 1H), 2.42-2.31 (m, 1H), 2.30-2.17 (m, 1H), 1.93-1.79 (m,1H), 1.75-1.64 (m, 1H), 1.50 (s, 9H), 1.30-1.23 (m, 6H)

Preparation of Compound 177

General method J, trans-tert-butyl2-(5-bromothiazol-2-yl)-5-(isopropoxycarbonylamino)piperidine-1-carboxylate. ESI [M+H]=450.2/448.2

Preparation of Compound 178

General method B, trans-tert-butyl2-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxycarbonylamino)phenyl]thiazol-2-yl]-5-(isopropoxycarbonylamino)piperidine-1-carboxylate.ESI [M+H]=682.3

Preparation of Ex. 63

General method C, trans-isopropylN-[6-[S-[2-(tert-butylsulfamoyl)-4-(isopropoxycarbonylamino)phenyl]thiazol-2-yl]-3-piperidyl]carbamate.¹H NMR (400 MHz, METHANOL-d4) δ=8.40 (d, J=2.0 Hz, 1H), 7.86 (s, 1H),7.65 (dd, J=2.2, 8.4 Hz, 1H), 7.38 (d, J=8.4 Hz, 1H), 5.05-4.94 (m, 1H),4.85-4.81 (m, 1H), 4.67 (dd, J=2.9, 11.9 Hz, 1H), 3.92-3.82 (m, 1H),3.60 (br dd, J=3.6, 11.8 Hz, 1H), 2.92 (t, J=11.9 Hz, 1H), 2.48 (br dd,J=3.2, 14.2 Hz, 1H), 2.22-2.03 (m, 2H), 1.82-1.70 (m, 1H), 1.32 (d,J=6.4 Hz, 6H), 1.24 (br d, J=6.2 Hz, 6H), 1.19 (s, 9H). ESI [M+H]=582.2

Example 64 Synthesis of isopropyl((3R,6S)-6-(5-(4-(3-benzylureido)-2-(N-(tert-butyl)sulfamoyl)phenyl)thiazol-2-yl)piperidin-3-yl)carbamate

The following compound was synthesized via same method by the keyintermediate 178.

¹H NMR (400 MHz, METHANOL-d4) δ=8.30 (d, J=2.2 Hz, 1H), 7.86 (s, 1H),7.66 (dd, J=2.2, 8.4 Hz, 1H), 7.38-7.20 (m, 6H), 4.85-4.82 (m, 1H), 4.70(dd, J=3.2, 12.0 Hz, 1H), 4.42 (s, 2H), 3.93-3.82 (m, 1H), 3.62 (br dd,J=3.5, 11.7 Hz, 1H), 3.00-2.90 (m, 1H), 2.52-2.44 (m, 1H), 2.23-2.04 (m,2H), 1.84-1.70 (m, 1H), 1.26-1.21 (m, 6H), 1.18 (s, 9H). ESI [M+H]=629.2

Example 65 Synthesis of isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[3-(isopropoxycarbonylamino)cyclobutyl]thiazol-5-yl]phenyl]carbamate

Preparation of Compound 180

To a solution of trans-3-(tert-butoxycarbonylamino)cyclobutanecarboxylicacid (500 mg, 2.32 mmol, 1 eq.), DIEA (750.54 mg, 5.81 mmol, 1.01 mL,2.5 eq.) in THF (10 mL) was added ETHYL CHLOROFORMATE (277.29 mg, 2.56mmol, 1.1 eq.) and the mixture was stirred 0° C. for 1 hr. Then it wasadded into NH₃.H₂O (1.30 g, 9.29 mmol, 1.43 mL, 25% purity, 4 eq.) inTHF (10 mL) and dioxane (10 mL) and the mixture was stirred at 25° C.for 1 hr. The mixture was washed with 1N HCl (20 mL), sat.aq.Na₂CO₃ (20mL) and the organic phase was dried over Na₂SO₄, filtered andconcentrated to give trans-tert-butyl N-(3-carbamoylcyclobutyl)carbamate (0.39 g, crude) as a white solid. ¹H NMR (400 MHz, DMSO-d6)δ=7.20-7.07 (m, 2H), 6.71 (br s, 1H), 4.09-4.00 (m, 1H), 2.72 (br t,J=9.3 Hz, 1H), 2.23 (br t, J=8.7 Hz, 2H), 2.01 (q, J=9.8 Hz, 2H), 1.34(s, 9H. ESI [M+Na]=237.1

Preparation of Compound 181

General method L, trans-tert-butylN-(3-carbamothioylcyclobutyl)carbamate. 1H NMR (400 MHz, DMSO-d6) δ=9.33(br s, 1H), 9.02 (br s, 1H), 7.18 (br d, J=5.5 Hz, 1H), 4.16-4.03 (m,1H), 3.33 (br s, 1H), 2.45 (br d, J=12.2 Hz, 2H), 2.15 (br d, J=6.5 Hz,2H), 1.37 (br s, 9H)

Preparation of Compound 182

General method M, trans-tert-butylN-(3-thiazol-2-ylcyclobutyl)carbamate. ¹H NMR (400 MHz, CHLOROFORM-d)δ=7.62 (d, J=3.2 Hz, 1H), 7.14 (dd, J=3.3, 8.3 Hz, 1H), 4.79 (br s, 1H),4.34 (br s, 1H), 3.47-3.36 (m, 1H), 2.81 (br d, J=8.8 Hz, 1H), 2.66(ddd, J=4.5, 7.9, 12.8 Hz, 1H), 2.46-2.34 (m, 1H), 2.17-2.09 (m, 1H),1.38 (s, 9H). ESI[M+H]=255.3

Preparation of Compound 183

General method J, trans-tert-butylN-[3-(5-bromothiazol-2-yl)cyclobutyl]carbamate. ESI[M+H]=335.1/333.1

Preparation of Compound 184

General method K, trans-tert-butylN-[3-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclobutyl]carbamate.ESI [M+H]=481.0

Preparation of Compound 185

General method F,trans-5-amino-2-[2-(3-aminocyclobutyl)thiazol-5-yl]-N-tert-butyl-benzenesulfonamide.ESI [M+H]=381.2

Preparation of Ex. 65

General method D, isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[3-(isopropoxycarbonylamino)cyclobutyl]thiazol-5-yl]phenyl]carbamate.¹H NMR (400 MHz, METHANOL-d4) δ=8.34 (d, J=1.5 Hz, 1H), 7.76-7.65 (m,2H), 7.38 (t, J=8.5 Hz, 1H), 5.04-4.94 (m, 1H), 4.82 (br d, J=6.4 Hz,1H), 4.42-4.32 (m, 0.5H), 4.13 (br t, J=8.0 Hz, 0.5H), 3.87-3.78 (m,0.5H), 3.58-3.48 (m, 0.5H), 2.82 (dq, J=2.6, 7.9 Hz, 1H), 2.72-2.64 (m,1H), 2.60-2.50 (m, 1H), 2.34-2.23 (m, 1H), 1.31 (d, J=6.2 Hz, 6H), 1.22(br d, J=6.0 Hz, 6H), 1.13 (s, 9H). ESI [M+H]=553.4

Examples 66A and 66B

Preparation of Compound 31.

To a solution of 2-bromobenzenesulfonyl chloride (100.00 g, 391.36 mmol,1.00 eq.) in H₂SO₄ (1.0 L) was added a solution of HNO₃ (79.95 g, 1.21mol, 57.11 mL, 95% purity, 3.08 eq.) in H₂SO₄ (0.50 L) drop-wise at 0°C. The mixture was stirred at 26° C. for 2 hrs. TLC (Petroleumether:EtOAc=10:1, R_(f)=0.40) showed the reaction was complete. Themixture was added slowly to ice water (5 L) with vigorous stirring andthen filtered. The filter cake was washed with H₂O (1 L×3) and dried togive 2-bromo-5-nitro-benzenesulfonyl chloride (105.00 g, crude) as ayellow solid. ¹H NMR (400 MHz, CHLOROFORM-d) δ=8.98 (d, J=2.4 Hz, 1H),8.37 (dd, J=2.5, 8.7 Hz, 1H), 8.09 (d, J=8.6 Hz, 1H).

Preparation of Compound 32.

To a mixture of 2-methylpropan-2-amine (200 g, 2.73 mol, 287.36 mL, 3.29eq.) and DMAP (10 g, 81.85 mmol, 0.1 eq.) in DCM (2 L) was added2-bromo-5-nitro-benzenesulfonyl chloride (250 g, 831.91 mmol, 1 eq.)portionwise at 0° C. The mixture was warmed to 15° C. and stirred for 1hr. LCMS showed the reaction was complete. The mixture was washed withHCl (1 N, 2 L), sat.aq. NaHCO₃ (500 mL) and brine (500 mL), dried overNa₂SO₄, filtered and concentrated to give2-bromo-N-tert-butyl-5-nitro-benzenesulfonamide (240 g, crude) as a graysolid. ESI [M+H]=336.9/338.9.

Preparation of Compound 33.

To a solution of 2-bromo-N-tert-butyl-5-nitro-benzenesulfonamide (35 g,103.80 mmol, 1 eq.) in DMA (300 mL), were added thiazole (26.51 g,311.40 mmol, 3 eq.), Pd(OAc)₂ (2.33 g, 10.38 mmol, 0.1 eq.) and KOAc(30.56 g, 311.40 mmol, 3 eq.). The mixture was stirred at 140° C. for 16hrs under N₂. LCMS showed the reaction was complete, the mixture waspoured into water (3 L) and filtered. The filter cake was washed withwater (200 mL×3) and then dried to giveN-tert-butyl-5-nitro-2-thiazol-5-yl-benzenesulfonamide (25.1 g, crude)as a black brown solid. ¹H NMR (400 MHz, DMSO-d6) δ=9.30 (s, 1H), 8.81(d, J=2.4 Hz, 1H), 8.46 (dd, J=2.4, 8.3 Hz, 1H), 8.12 (s, 1H), 7.83 (d,J=8.3 Hz, 1H), 7.60 (s, 1H), 1.07 (s, 9H). ESI [M+H]=342.0.

Preparation of Compound 34.

To a solution of N-tert-butyl-5-nitro-2-thiazol-5-yl-benzenesulfonamide(15 g, 43.94 mmol, 1 eq.) in AcOH (200 mL), were added KOAc (21.56 g,219.68 mmol, 5 eq.) and Br₂ (35.11 g, 219.68 mmol, 5 eq.). The mixturewas stirred at 80° C. for 3 hrs. LCMS showed the reaction was complete,the mixture was quenched by sat.aq.Na₂CO₃ (1 L) and extracted with EtOAc(300 mL×3). The combined organic phase was dried over Na₂SO₄, filteredand concentrated to give2-(2-bromothiazol-5-yl)-N-tert-butyl-5-nitro-benzenesulfonamide (18 g,crude) as a green solid which was used without any purification.

Preparation of Compound 35.

To a solution of2-(2-bromothiazol-5-yl)-N-tert-butyl-5-nitro-benzenesulfonamide (8.5 g,20.22 mmol, 1 eq.) in EtOH (70 mL), THF (40 mL) and H₂O (20 mL), wereadded Fe (3.39 g, 60.67 mmol, 3 eq.) and NH₄Cl (3.25 g, 60.67 mmol, 2.12mL, 3 eq.). The mixture was stirred at 90° C. for 1 hr. LCMS showed thereaction was complete. The mixture was filtered, the filtrate wasconcentrated to remove organic solvent and extracted with DCM (200mL×2). The organic phase was dried over Na₂SO₄, filtered andconcentrated to afford5-amino-2-(2-bromothiazol-5-yl)-N-tert-butyl-benzenesulfonamide (6 g,crude) as a yellow solid. ¹HNMR showed the structure was correct. 1H NMR(400 MHz, DMSO-d6) δ=7.58-7.52 (m, 1H), 7.30 (d, J=2.4 Hz, 1H), 7.11 (d,J=8.3 Hz, 1H), 6.99 (s, 1H), 6.73 (dd, J=2.4, 8.3 Hz, 1H), 5.94 (s, 2H),1.07 (s, 9H). ESI [M+H]=390.0/392.0.

Preparation of Compound 36.

General method D,(4-nitrophenyl)N-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl)phenyl]carbamateas DCM solution ESI [M+H]=555.0/557.0

Preparation of Compound 37.

General method H,1-benzyl-3-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl) phenyl]urea.ESI [M+H]=523.0/525.0.

Preparation of Compound 38.

General method B (Suzuki reaction),tert-butyl-N-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohex-3-en-1-yl]carbamate(Compound 38)

To a solution of Compound 17 (1 eq.) in dioxane and H₂O, were addedPd(dppf)Cl₂ (0.1 eq.), Compound 37 (0.9 eq.) and Na₂CO₃ (3 eq.). Themixture was stirred at 80° C. for 12 hrs under N₂. LCMS showed thereaction was complete. The mixture was concentrated and the residue waspurified by prep-TLC (PE/EtOAc=1:1) to yield product 38. ESI [M+H]=640.5

Preparation of Compound 39.

tert-butylN-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate

To a solution of Compound 38 (1 eq.) and AcOH (0.1 eq.) in EtOAc wasadded Pd/C (10% purity, 1.00 eq.). The mixture was stirred under H₂ (15psi) at 40° C. for 1 hr. LCMS showed the reaction was complete and thenthe mixture was filtered and concentrated. The residue was purified bycolumn chromatography (SiO₂, Petroleum ether/Ethyl acetate=10/1 to 1:1)to give Compound 39. ESI [M+H]=642.5

Preparation of Compound 40.

General method F,1-[4-[2-(4-aminocyclohexyl)thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]-3-benzyl-urea. ESI [M+H]=542.5.

Preparation of Compound 41

General method D, isopropylN-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate,ESI [M+H]=628.3.

Preparation of Compound Ex. 66A and Ex. 66B.

Compound 41 was separated by SFC (Instrument: Thar SFC80 preparativeSFC; Column: Chiralpak AD-H 250*30 mm i.d. 5u; Mobile phase: A for CO₂and B for IPA (0.1% NH₃H₂O); Gradient: B %=42%; Flow rate: 70 g/min;Wavelength: 220 nm; Column temperature: 40° C.; System back pressure:100 bar; Cycle time: 20 min; Injection amount: 4 mg per injection), andthen purified by prep-HPLC (Column: Luna C18 100*30 5u; mobile phase:[water(0.1% TFA)-ACN]; B %: 35%-75%,5 min). trans-isopropylN-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamateEx. 66A (11.54 mg, 18.38 umol, 5.53% yield, 100% purity) andcis-isopropylN-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate Ex. 66B (9.38 mg, 14.94 umol, 4.50%yield, 100% purity) were obtained.

Trans-isopropylN-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate

¹H NMR (400 MHz, METHANOL-d4) δ=8.24 (d, J=2.2 Hz, 1H), 7.72 (s, 1H),7.69 (dd, J=2.6, 8.3 Hz, 1H), 7.37-7.30 (m, 5H), 7.27-7.22 (m, 1H),4.85-4.78 (m, 1H), 4.41 (s, 2H), 3.50-3.39 (m, 1H), 3.06-2.96 (m, 1H),2.22 (br d, J=11.8 Hz, 2H), 2.07 (br d, J=10.1 Hz, 2H), 1.76-1.62 (m,2H), 1.41 (dq, J=3.1, 12.7 Hz, 2H), 1.22 (br d, J=6.1 Hz, 6H), 1.11 (s,9H). ESI [M+H]=628.2.

Cis-isopropylN-[4-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate

¹H NMR (400 MHz, METHANOL-d4) δ=8.25 (d, J=2.2 Hz, 1H), 7.74 (s, 1H),7.70 (dd, J=2.2, 8.3 Hz, 1H), 7.38-7.30 (m, 5H), 7.28-7.22 (m, 1H), 4.82(br d, J=6.1 Hz, 1H), 4.41 (s, 2H), 3.74 (br s, 1H), 3.20-3.13 (m, 1H),2.03-1.95 (m, 4H), 1.86-1.72 (m, 4H), 1.23 (d, J=6.1 Hz, 6H), 1.12 (s,9H). ESI [M+H]=628.2.

Examples 67A and 67B

Preparation of Compound 42.

General method D, isopropylN-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl) phenyl]carbamate. ESI[M+H]=476.0/478.0.

Preparation of Compound 43

General method B, isopropylN-[4-[2-[4-(tert-butoxycarbonylamino)cyclohexen-1-yl]thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]carbamate.ESI [M+H]=593.3.

Preparation of Compound 44.

General method I, isopropylN-[4-[2-[4-(tert-butoxycarbonylamino)cyclohexyl]thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]carbamate. ESI [M+H]=595.3.

Preparation of Compound 45.

General method F, isopropyl N-[4-[2-(4-aminocyclohexyl)thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]carbamate. ESI [M+H]=495.2.

Preparation of Compound 46.

General method D, isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate. ESI [M+H]=581.2.

Preparation of Ex. 67A and Ex. 67B.

Compound 46 was separated by SFC (Instrument: Thar SFC80 preparativeSFC; Column: ChiralpakAD-H 250*30 mm i.d. 5u; Mobile phase: A for CO₂and B for IPA(0.1% NH₃H₂O); Gradient: B %=30%; Flow rate:70 g/min;Wavelength:220 nm; Column temperature: 40° C.; System back pressure: 100bar; Cycle time:8 min; Injection amount: 3 mg per injection); and thenpurified by prep-HPLC (Column: Agela Durashell C18 150*25 5u; mobilephase: [water(0.1% TFA)-ACN]; B %: 55%-85%,12 min),trans-isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamateEx. 67A (5.76 mg, 100% purity) andcis-isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate Ex. 67B (3.95 mg, 100% purity)were obtained as a pale yellow solid.

Trans-isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate(Compound S12)

¹H NMR (400 MHz, METHANOL-d4) δ=8.37 (d, J=2.3 Hz, 1H), 7.75 (s, 1H),7.69 (dd, J=2.2, 8.4 Hz, 1H), 7.40 (d, J=8.3 Hz, 1H), 5.01 (td, J=6.3,12.5 Hz, 1H), 4.86-4.82 (m, 1H), 3.52-3.42 (m, 1H), 3.04 (tt, J=3.5,12.0 Hz, 1H), 2.35-2.19 (m, 2H), 2.15-1.99 (m, 2H), 1.72 (dq, J=3.0,12.9 Hz, 2H), 1.43 (dq, J=3.3, 12.6 Hz, 2H), 1.34 (d, J=6.2 Hz, 6H),1.25 (br d, J=6.1 Hz, 6H), 1.14 (s, 9H). ESI [M+H]=581.2.

Cis-isopropylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate(Compound S13)

¹H NMR (400 MHz, METHANOL-d4) δ=8.37 (d, J=2.2 Hz, 1H), 7.76 (s, 1H),7.70 (dd, J=2.2, 8.3 Hz, 1H), 7.40 (d, J=8.3 Hz, 1H), 5.01 (td, J=6.3,12.5 Hz, 1H), 4.84 (br s, 1H), 3.77 (br s, 1H), 3.23-3.15 (m, 1H),2.05-1.98 (m, 4H), 1.89-1.72 (m, 4H), 1.34 (d, J=6.2 Hz, 6H), 1.25 (d,J=6.1 Hz, 6H), 1.16 (s, 9H). ESI [M+H]=581.2

Example 68 Synthesis of isopropylN-[1-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-4-piperidyl]carbamate

Preparation of Compound 24.

A mixture of1-benzyl-3-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl)phenyl]urea(0.06 g, 114.62 umol, 1.0 eq.), tert-butyl N-(4-piperidyl)carbamate (40mg, 199.72 umol, 1.74 eq.), Xantphos (6.63 mg, 11.46 umol, 0.1 eq.),Pd(OAc)₂ (2.57 mg, 11.46 umol, 0.1 eq.) and Cs₂CO₃ (112.04 mg, 343.86umol, 3.0 eq.) in ACN (3 mL) was heated to 80° C. for 12 hrs under N₂.LCMS showed the reaction was complete and the mixture was concentrated.The residue was purified by prep-TLC (Petroleum ether:Ethyl acetate=1:1)to give tert-butylN-[1-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-4-piperidyl]carbamate(0.03 g, crude) as a yellow solid. ESI [M+H]=643.1.

Preparation of Compound 25.

Tert-butylN-[1-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-4-piperidyl]carbamate

(30 mg, 46.67 umol, 1.0 eq.) was dissolved into HCl/MeOH (4M, 1 mL) andthe mixture was stirred at 20° C. for 0.5 hr. LCMS showed the reactionwas complete and the mixture was concentrated to give1-[4-[2-(4-amino-1-piperidyl)thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]-3-benzyl-urea(27 mg, crude, HCl salt) as a yellow solid. ESI [M+H]=543.2.

Preparation of Ex. 68.

General method D, isopropylN-[1-[5-[4-(benzylcarbamoylamino)-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]-4-piperidyl]carbamate. ¹H NMR (400 MHz,METHANOL-d4) δ=8.24 (d, J=2.2 Hz, 1H), 7.69 (dd, J=2.4, 8.4 Hz, 1H),7.39 (d, J=8.4 Hz, 1H), 7.36-7.19 (m, 6H), 4.82-4.73 (m, 1H), 4.40 (s,2H), 3.92 (br d, J=13.7 Hz, 2H), 3.75-3.68 (m, 1H), 3.43 (br t, J=11.0Hz, 2H), 2.10-2.02 (m, 2H), 1.71-1.60 (m, 2H), 1.22 (br d, J=6.2 Hz,6H), 1.18 (s, 9H). ESI [M+H]=629.2.

Example 69 Synthesis of isopropylN-[1-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxycarbonylamino)phenyl]thiazol-2-yl]-4-piperidyl]carbamate

Preparation of Compound 26.

General method D, isopropylN-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl) phenyl]carbamate. ESI[M+H]=478.0/476.0.

Preparation of Compound 27.

To a solution of isopropylN-[4-(2-bromothiazol-5-yl)-3-(tert-butylsulfamoyl) phenyl]carbamate(0.05 g, 104.95 umol, 1.0 eq.) in MeCN (2 mL) were added Cs₂CO₃ (102.59mg, 314.86 umol, 3.0 eq.), KI (17.42 mg, 104.95 umol, 1.0 eq.) andtert-butyl N-(4-piperidyl)carbamate (105.10 mg, 524.76 umol, 5.0 eq.).The mixture was stirred at 95° C. for 12 hrs and then concentrated. Theresidue was purified by prep-TLC (SiO₂, Petroleum ether:Ethylacetate=4:3) to giveN-[4-[2-[4-(tert-butoxycarbonylamino)-1-piperidyl]thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]carbamate (0.025 g, crude) as a white solid. ESI [M+H]=596.2.

Preparation of Compound 28.

IsopropylN-[4-[2-[4-(tert-butoxycarbonylamino)-1-piperidyl]thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]carbamate

(0.025 g, 41.96 umol, 1.0 eq.) was dissolved into HCl/MeOH (4M, 1.5 mL)and the mixture was stirred at 20° C. for 0.5 hr. LCMS showed thereaction was complete and the mixture was concentrated to give theN-[4-[2-(4-amino-1-piperidyl)thiazol-5-yl]-3-(tert-butylsulfamoyl)phenyl]carbamate (0.02 g, crude, HCl salt) as a white solid. ESI [M+H]=496.2.

Preparation of Ex. 69.

General method D, isopropylN-[1-[5-[2-(tert-butylsulfamoyl)-4-(isopropoxycarbonylamino)phenyl]thiazol-2-yl]-4-piperidyl]carbamate. ¹H NMR (400MHz, METHANOL-d4) δ=8.35 (s, 1H), 7.66 (br d, J=8.4 Hz, 1H), 7.42 (d,J=8.4 Hz, 1H), 7.31 (s, 1H), 4.98 (td, J=6.2, 12.5 Hz, 1H), 4.84-4.77(m, 1H), 3.93 (br d, J=13.5 Hz, 2H), 3.77-3.67 (m, 1H), 3.44 (br t,J=12.3 Hz, 2H), 2.06 (br dd, J=3.1, 13.2 Hz, 2H), 1.71-1.61 (m, 2H),1.31 (d, J=6.4 Hz, 6H), 1.23 (br d, J=6.2 Hz, 6H), 1.19 (s, 9H). ESI[M+H]=582.2.

Example 70 Synthesis of trans-isopropylN-[4-[5-[2-(tert-butylsulfamoyl)-4-[(2-fluorophenyl)methoxycarbonylamino]phenyl]thiazol-2-yl]cyclohexyl]carbamate

Preparation of Compound 2.

To a mixture of trans-4-(tert-butoxycarbonylamino)cyclohexanecarboxylicacid (65.0 g, 267.2 mmol, 1.0 eq.), NH₄Cl (21.4 g, 400.7 mmol, 1.5 eq.)and TEA (801.5 mmol, 111.6 mL, 3 eq.) in MeCN (1.3 L) was added HBTU(111.5 g, 293.9 mmol, 1.1 eq.) and the mixture was stirred at 25° C. for3 hrs. The mixture was filtered and then the filter cake was washed withpetroleum ether (200 mL) and dried to give trans-tert-butylN-(4-carbamoylcyclohexyl)carbamate (140 g, crude, 2 batches) as a whitesolid. ¹H NMR (METHANOL-d4, 400 MHz) δ=3.25-3.34 (m, 1H), 2.14 (tt,J=12.3, 3.5 Hz, 1H), 1.83-1.99 (m, 4H), 1.52 (qd, J=13.1, 2.9 Hz, 2H),1.42 (s, 9H), 1.21 (qd, J=12.7, 3.5 Hz, 2H)

Preparation of Compound 3.

A mixture of trans-tert-butyl N-(4-carbamoylcyclohexyl)carbamate (90.0g, 371.4 mmol, 1.0 eq.), Na₂CO₃ (39.4 g, 371.4 mmol, 1.0 eq.) andLawesson's reagent (82.6 g, 204.3 mmol, 0.55 eq.) in 2-Me-THF (600 mL)was stirred at 80° C. for 2 hrs and then the reaction mixture was pouredinto H₂O (200 mL). The aqueous phase was extracted with EtOAc (500mL×2). The combined organic layers were dried over Na₂SO₄, filtered andconcentrated to give trans-tert-butylN-(4-carbamothioylcyclohexyl)carbamate (180 g, crude, 2 batches) as awhite solid. ¹H NMR (METHANOL-d4, 400 MHz) δ=3.35-3.46 (m, 1H),2.87-3.00 (m, 1H), 2.09-2.20 (m, 2H), 1.99-2.09 (m, 2H), 1.54-1.68 (m,2H), 1.26-1.45 (m, 2H), 1.14-1.25 (m, 9H)

Preparation of Compound 4.

A mixture of trans-tert-butyl N-(4-carbamothioylcyclohexyl)carbamate(180.0 g, 696.7 mmol, 1.0 eq.), 2-bromo-1,1-diethoxy-ethane (137.3 g,696.7 mmol, 1.0 eq.) and TsOH.H₂O (265 g, 1.4 mol, 2 eq.) in EtOH (2.0L) was stirred at 80° C. for 6 hrs. Then the mixture was cooled to RTand adjusted to PH=9 with aq.sat.Na₂CO₃ and Boc₂O (152 g, 696.7 mmol, 1eq.) was added. The mixture was stirred at 30° C. for 3 hrs, thenconcentrated and diluted with H₂O (2 L). The mixture was extracted withEtOAc (800 mL×3) and the combined organic layers were dried over Na₂SO₄,filtered and concentrated. The residue was triturated with petroleumether (1.5 L) to give trans-tert-butyl N-(4-thiazol-2-ylcyclohexyl)carbamate (70 g, 247.87 mmol, 35.58% yield) as a white solid. ¹H NMR(METHANOL-d4, 400 MHz) δ=7.67 (d, J=3.1 Hz, 1H), 7.44 (d, J=3.5 Hz, 1H),3.33-3.44 (m, 1H), 2.93-3.06 (m, 1H), 2.12-2.21 (m, 2H), 2.00-2.09 (m,2H), 1.57-1.71 (m, 2H), 1.41-1.48 (m, 9H), 1.29-1.38 (m, 1H), 1.13-1.28(m, 1H).

Preparation of Compound 5.

A mixture of trans-tert-butyl N-(4-thiazol-2-ylcyclohexyl)carbamate (68g, 240.8 mmol, 1 eq.) and NBS (47.1 g, 264.9 mmol, 1.1 eq.) in DMF (500mL) was stirred at 25° C. for 10 hrs and then poured into H₂O (2 L) andextracted with EtOAc (500 mL×3). The combined organic layers were washedwith brine (300 mL×5), dried over Na₂SO₄, filtered and concentrated. Theresidue was purified by column chromatography (SiO₂, Petroleumether:Ethyl acetate=20:1 to 10:1) to give trans-tert-butylN-[4-(5-bromothiazol-2-yl)cyclohexyl]carbamate (78 g, crude) as a yellowsolid. ESI [M+H]=363.0/361.0

Preparation of Compound 6.

A mixture of trans-tert-butylN-[4-(5-bromothiazol-2-yl)cyclohexyl]carbamate (50 g, 138.4 mmol, 1 eq.)in HCl/MeOH (4 M, 700 mL) was stirred at 25° C. for 0.5 hr and thenconcentrated to give trans-4-(5-bromothiazol-2-yl) cyclohexanamine (45g, crude, HCl salt) as a yellow solid. ESI [M+H]=263.0/261.0

Preparation of Compound 7.

To a solution of trans-4-(5-bromothiazol-2-yl)cyclohexanamine (45 g, HClsalt, 172.3 mmol, 1 eq.), Pyridine (61.5 mmol, 69.5 mL, 5 eq.) and DMAP(4.2 g, 34.5 mmol, 0.2 eq.) in DCM (300 mL) was added isopropylcarbonochloridate (258.5 mmol, 35.9 mL, 1.5 eq.) dropwise at 0° C. Themixture was stirred at 25° C. for 0.5 hr and then washed with HCl (1N, 1L) and sat.aq.Na₂CO₃ (1 L). The organic layers were dried over Na₂SO₄,filtered and concentrated. The residue was purified by columnchromatography (SiO₂, Petroleum ether:Ethyl acetate=15:1 to 10:1) togive trans-isopropyl N-[4-(5-bromothiazol-2-yl)cyclohexyl]carbamate (37g, 106.55 mmol, 61.84% yield) as a yellow solid. ¹H NMR (METHANOL-d4,400 MHz) δ=7.60 (s, 1H), 4.81 (dt, J=12.2, 6.2 Hz, 1H), 3.41 (tt,J=11.6, 4.0 Hz, 1H), 2.88-3.00 (m, 1H), 2.10-2.20 (m, 2H), 1.98-2.07 (m,2H), 1.55-1.68 (m, 2H), 1.30-1.43 (m, 2H), 1.21 (br d, J=6.2 Hz, 6H).

Preparation of Compound 8.

A mixture of trans-isopropylN-[4-(5-bromothiazol-2-yl)cyclohexyl]carbamate (16.2 g, 46.7 mmol, 1eq.),5-amino-N-tert-butyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide(19.9 g, 56.1 mmol, 1.2 eq.), KF (4.1 g, 70.1 mmol, 1.5 eq.), Na₂CO₃(14.9 g, 140.2 mmol, 3 eq.) and Pd(PPh₃)₄(1.6 g, 1.4 mmol, 0.03 eq.) intoluene (150 mL), EtOH (150 mL) and H₂O (50 mL) stirred at 80° C. for 6hrs under N₂ atmosphere. The reaction mixture was concentrated and theresidue was diluted with H₂O (100 mL) and extracted with EtOAc (100mL×2). The combined organic layers were dried over Na₂SO₄, filtered andconcentrated. The residue was purified by column chromatography (SiO₂,Petroleum ether:Ethyl acetate=20:1 to 1:1) to give trans-isopropylN-[4-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate (13 g, 26.3 mmol, 56.2% yield)as a white solid. ¹H NMR (400 MHz, METHANOL-d4) δ=7.65-7.60 (m, 1H),7.43 (d, J=2.2 Hz, 1H), 7.13 (d, J=8.3 Hz, 1H), 6.83 (dd, J=2.6, 8.3 Hz,1H), 4.81 (br s, 1H), 3.76-3.71 (m, 1H), 3.05-2.86 (m, 1H), 2.20 (br d,J=12.3 Hz, 2H), 2.06 (br d, J=10.5 Hz, 2H), 1.75-1.60 (m, 2H), 1.47-1.34(m, 2H), 1.22 (br d, J=6.1 Hz, 6H), 1.09 (s, 9H).

Preparation of Compound 9.

To a solution of trans-isopropylN-[4-[5-[4-amino-2-(tert-butylsulfamoyl)phenyl]thiazol-2-yl]cyclohexyl]carbamate (250 mg, 505.4 umol, 1 eq.) in DCM (4mL) were added DMAP (6.2 mg, 50.5 umol, 0.1 eq.), Pyridine (120 mg, 1.5mmol, 3 eq.) and (4-nitrophenyl) carbonochloridate (153 mg, 758 umol,1.5 eq.). The mixture was stirred at 25° C. for 0.5 hr and used directlyfor the next step. ESI [M+H]=660.2

Preparation of Compound Ex. 70.

To a solution of (2-fluorophenyl)methanol (45.9 mg, 363.8 umol, 3 eq.)and DIEA (47 mg, 363.8 umol, 3 eq.) in MeCN (1 mL) was added the abovedsolution (1 mL). The mixture was stirred at 80° C. for 1 hr, thenconcentrated and the residue was purified by prep-HPLC (Column: WatersXbridge 150*25 5u; Mobile phase: [water (10 mM NH₄HCO₃)-ACN]; B %:42%-72%, 12 min) to give trans-isopropylN-[4-[5-[2-(tert-butylsulfamoyl)-4-[(2-fluorophenyl)methoxycarbonylamino]phenyl]thiazol-2-yl]cyclohexyl]carbamate (16.34 mg, 25.26umol, 20.83% yield, 100% purity) as a yellow solid. ¹H NMR (400 MHz,METHANOL-d4) δ=8.36 (s, 1H), 7.77-7.63 (m, 2H), 7.52 (t, J=7.0 Hz, 1H),7.44-7.31 (m, 2H), 7.23-7.08 (m, 2H), 5.29 (s, 2H), 4.83 (br s, 1H),3.46 (br d, J=11.8 Hz, 1H), 3.00 (br t, J=11.8 Hz, 1H), 2.22 (br d,J=12.7 Hz, 2H), 2.07 (br d, J=11.4 Hz, 2H), 1.76-1.62 (m, 2H), 1.47-1.35(m, 2H), 1.22 (br d, J=6.1 Hz, 6H), 1.12 (s, 9H). ESI [M+H]=647.2

Example 71 Synthesis of trans-[(1S)-1-phenylethyl]N-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate

Preparation of Ex. 71.

To a solution of (1S)-1-phenylethanol (29.6 mg, 242.5 umol, 2 eq.) andDIEA (47 mg, 363.8 umol, 3 eq.) in MeCN (2 mL) was added a solution oftrans-(4-nitrophenyl)N-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate(80 mg, 121.25 umol, 1 eq.) in DCM (1 mL). The mixture was stirred at80° C. for 1 hr, then concentrated and the residue was purified byprep-HPLC (Column: Nano-Micro UniSil 5-100 C18 ULTRA 100*250 mm 5 um;mobile phase: [water(0.1% TFA)-ACN]; B %: 55%-80%,11 min) to givetrans-[(1S)-1-phenylethyl]N-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate(12.19 mg, 18.77 umol, 15.48% yield, 99% purity) as a white solid. ¹HNMR (400 MHz, METHANOL-d4) δ=8.35 (br s, 1H), 7.81-7.65 (m, 2H),7.48-7.36 (m, 5H), 7.31 (br d, J=6.8 Hz, 1H), 5.89 (br d, J=6.2 Hz, 1H),4.85 (br d, J=5.5 Hz, 1H), 3.47 (br s, 1H), 3.03 (br s, 1H), 2.24 (br d,J=11.2 Hz, 2H), 2.08 (br d, J=11.0 Hz, 2H), 1.71 (q, J=11.9 Hz, 2H),1.61 (br d, J=6.4 Hz, 3H), 1.49-1.37 (m, 2H), 1.24 (br d, J=5.4 Hz, 6H),1.13 (s, 9H). ESI [M+H]=643.2

Example 72 Synthesis ofN-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate

Preparation of Ex. 72.

To a solution of 2-pyridylmethanol (26.5 mg, 242.5 umol, 2 eq.) and DIEA(47 mg, 363.8umol, 3 eq.) in MeCN (2 mL) was added a solution oftrans-(4-nitrophenyl)N-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate(80 mg, 121.3 umol, 1 eq.) in DCM (1 mL). The mixture was stirred at 80°C. for 1 hr, then concentrated and the residue was purified by prep-HPLC(Column: Waters Xbridge 150*25 5u; Mobile phase: [water (0.04% NH₃H₂O+10mM NH₄HCO₃)-ACN]; B %: 35%-65%, 10 min) to give trans-2-pyridylmethylN-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate (18.59 mg, 28.82 umol, 23.77% yield,97.65% purity) as a pale yellow solid. ¹H NMR (400 MHz, METHANOL-d4)δ=8.56 (br d, J=4.5 Hz, 1H), 8.38 (d, J=1.8 Hz, 1H), 7.94-7.87 (m, 1H),7.78-7.72 (m, 2H), 7.59 (br d, J=7.8 Hz, 1H), 7.44-7.38 (m, 2H), 5.32(s, 2H), 4.85 (td, J=5.9, 12.0 Hz, 1H), 3.47 (br t, J=11.8 Hz, 1H),3.07-2.96 (m, 1H), 2.24 (br d, J=12.3 Hz, 2H), 2.13-2.04 (m, 2H),1.77-1.65 (m, 2H), 1.48-1.37 (m, 2H), 1.24 (br d, J=6.1 Hz, 6H), 1.14(s, 9H). ESI [M+H]=630.2

Example 73 Synthesis of trans-isopropylN-[4-[5-[2-(tert-butylsulfamoyl)-4-[(4-hydroxyphenyl)methylcarbamoylamino]phenyl]thiazol-2-yl]cyclohexyl]carbamate

Preparation of Ex. 73.

To a solution 4-(aminomethyl) phenol (74.7 mg, 606.5 umol, 3 eq.) andDIEA (78.4 mg, 606.5 umol, 3 eq.) in DCM (2 mL) was addedtrans-(4-nitrophenyl)N-[3-(tert-butylsulfamoyl)-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate(133.4 mg, 202.2 umol, 1 eq.) in DCM (2 mL). The mixture was stirred at25° C. for 1 hr, then concentrated and the residue was purified byprep-HPLC (TFA condition) to give trans-isopropylN-[4-[5-[2-(tert-butylsulfamoyl)-4-[(4-hydroxyphenyl)methylcarbamoylamino]phenyl]thiazol-2-yl]cyclohexyl]carbamate(23.56 mg, 35.99 umol, 17.80% yield, 98.357% purity) as a white solid.¹H NMR (400 MHz, METHANOL-d4) δ=8.24 (d, J=2.2 Hz, 1H), 7.77 (s, 1H),7.69 (dd, J=2.2, 8.3 Hz, 1H), 7.36 (d, J=8.3 Hz, 1H), 7.16 (d, J=8.8 Hz,2H), 6.75 (d, J=8.3 Hz, 2H), 4.85-4.74 (m, 1H), 4.30 (s, 2H), 3.45 (s,1H), 3.15-2.97 (m, 1H), 2.23 (br d, J=12.3 Hz, 2H), 2.08 (br d, J=13.2Hz, 2H), 1.79-1.63 (m, 2H), 1.48-1.34 (m, 2H), 1.22 (br d, J=6.1 Hz,6H), 1.12 (s, 9H). ESI [M+H]=644.2

Example 74 Synthesis of trans-isopropylN-[4-[5-[4-(benzylcarbamoylamino)-2-[(2-hydroxy-1,1-dimethyl-ethyl)sulfamoyl]phenyl]thiazol-2-yl]cyclohexyl]carbamate

Preparation of Compound 11.

To a mixture of 2-amino-2-methyl-propan-1-ol (3 g, 33.7 mmol, 5.1 eq.)and DMAP (80 mg, 654.8 umol, 0.98 eq.) in DCM (50 mL) was added2-bromo-5-nitro-benzenesulfonyl chloride (2 g, 6.66 mmol, 1 eq.). Themixture was stirred at 20° C. for 30 mins, then washed with 1N HCl (20mL) and sat.aq.NaHCO₃ (20 mL). The organic layer was dried over Na₂SO₄,filtered and concentrated to give2-bromo-N-(2-hydroxy-1,1-dimethyl-ethyl)-5-nitro-benzenesulfonamide (1.7g, 4.8 mmol, 72.3% yield) as a yellow gum. ESI [M+H]=355.0/353.0

Preparation of Compound 12.

A mixture of2-bromo-N-(2-hydroxy-1,1-dimethyl-ethyl)-S-nitro-benzenesulfonamide (1.7g, 4.8 mmol, 1 eq.), Fe (1.5 g, 26.9 mmol, 5.6 eq.) and NH₄Cl (800 mg,14.9 mmol, 3.1 eq.) in EtOH (15 mL)/H₂O (7.5 mL)/THF (7.5 mL) wasstirred at 80° C. for 2 hrs. The reaction mixture was concentrated toremove EtOH, diluted with H₂O (100 mL) and extracted with EtOAc (100mL×3). The combined organic layers were dried over Na₂SO₄, filtered andconcentrated. The residue was purified by column chromatography (SiO₂,Petroleum ether:Ethyl acetate=100:1 to 1:1) to give5-amino-2-bromo-N-(2-hydroxy-1,1-dimethyl-ethyl)benzenesulfonamide (1.1g, 3.4 mmol, 70.7% yield) as a pale yellow solid. ESI [M+H]=325.0/323.0

Preparation of Compound 13.

A mixture of 5-amino-2-bromo-N-(2-hydroxy-1,1-dimethyl-ethyl)benzenesulfonamide (400 mg, 1.24 mmol, 1 eq.),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(942.8 mg, 3.7 mmol, 3 eq.), Pd(dppf)Cl₂ (90.6 mg, 123.8 umol, 0.1 eq.)and KOAc (364.4 mg, 3.7 mmol, 3 eq.) in dioxane (4 mL) was stirred at80° C. for 12 hrs under N₂ atmosphere and then concentrated. The residuewas purified by column chromatography (SiO₂, Petroleum ether:Ethylacetate=50:1 to 3:1) to give5-amino-N-(2-hydroxy-1,1-dimethyl-ethyl)-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide(201 mg, crude) as a white solid. ¹H NMR (400 MHz, METHANOL-d4) δ=7.53(d, J=8.0 Hz, 1H), 7.28 (d, J=2.4 Hz, 1H), 6.75 (dd, J=6.4 Hz, 1H), 3.35(s, 2H), 1.36 (s, 12H), 1.10 (s, 6H).

Preparation of Compound 14.

A mixture of5-amino-N-(2-hydroxy-1,1-dimethyl-ethyl)-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzenesulfonamide(140.7 mg, 380.1 umol, 1.2 eq.), trans-isopropylN-[4-(5-bromothiazol-2-yl)cyclohexyl]carbamate (110 mg, 316.7 umol, 1eq.), Na₂CO₃ (100.7 mg, 950.3 umol, 3 eq.), KF (27.6 mg, 475.1 umol, 1.5eq.) and Pd(PPh₃)₄(36.6 mg, 31.7 umol, 0.1 eq.) in toluene(1 mL)/EtOH (1mL)/H₂O (0.3 mL) was stirred at 80° C. for 12 hrs under N₂ atmosphere.The reaction mixture was concentrated, diluted with H₂O (10 mL) andextracted with EtOAc (10 mL×2). The combined organic layers were driedover Na₂SO₄, filtered and concentrated. The residue was purified byprep-TLC (SiO₂, Ethyl acetate) to give trans-isopropylN-[4-[5-[4-amino-2-[(2-hydroxy-1,1-dimethyl-ethyl)sulfamoyl]phenyl]thiazol-2-yl]cyclohexyl]carbamate(117 mg, crude) as a yellow solid. ¹H NMR (400 MHz, METHANOL-d4) δ=7.63(d, J=7.0 Hz, 1H), 7.44 (d, J=2.2 Hz, 1H), 7.15 (d, J=8.3 Hz, 1H), 6.83(dd, J=2.4, 8.1 Hz, 1H), 4.85-4.78 (m, 1H), 3.50-3.39 (m, 1H), 3.25 (s,2H), 3.03-2.91 (m, 1H), 2.21 (br t, J=6.1 Hz, 2H), 2.12-2.00 (m, 2H),1.75-1.60 (m, 2H), 1.45-1.33 (m, 2H), 1.28-1.19 (s, 6H), 1.03 (s, 6H)

Preparation of Compound 15.

To a solution of trans-isopropylN-[4-[5-[4-amino-2-[(2-hydroxy-1,1-dimethyl-ethyl)sulfamoyl]phenyl]thiazol-2-yl]cyclohexyl]carbamate (90 mg, 176.3 umol, 1eq.) in DCM (1 mL) were added TEA (53.5 mg, 528.7 umol, 3 eq.), DMAP(2.2 mg, 17.6 umol, 0.1 eq.) and TBSCl (66.4 mg, 440.6 umol, 2.5 eq.).The mixture was stirred at 30° C. for 12 hrs and then concentrated. Theresidue was purified by prep-TLC (SiO₂, Petroleum ether:Ethylacetate=2:1) to give trans-isopropylN-[4-[5-[4-amino-2-[[2-[tert-butyl(dimethyl)silyl]oxy-1,1-dimethyl-ethyl]sulfamoyl]phenyl]thiazol-2-yl]cyclohexyl]carbamate(120 mg, crude) as a pale yellow solid. ESI [M+H]=625.2

Preparation of Compound 16.

To a solution of trans-isopropylN-[4-[5-[4-amino-2-[[2-[tert-butyl(dimethyl)silyl]oxy-1,1-dimethyl-ethyl]sulfamoyl]phenyl]thiazol-2-yl]cyclohexyl]carbamate(90 mg, 144 umol, 1 eq.) in DCM (1 mL) were added DMAP (1.8 mg, 14.4umol, 0.1 eq.), pyridine (34.2 mg, 432 umol, 3 eq.) and (4-nitrophenyl)carbonochloridate (43.5 mg, 216 umol, 1.5 eq.). The mixture was stirredat 25° C. for 0.5 hr and used into the next step directly withoutfurther purification. ESI [M+H]=790.3

Preparation of Compound 17.

To a solution of phenylmethanamine (36 mg, 336 umol, 3 eq.) in DCM (1mL) was added a solution oftrans-(4-nitrophenyl)N-[3-[[2-[tert-butyl(dimethyl)silyl]oxy-1,1-dimethyl-ethyl]sulfamoyl]-4-[2-[4-(isopropoxycarbonylamino)cyclohexyl]thiazol-5-yl]phenyl]carbamate(88.5 mg, 112 umol, 1 eq.) in DCM (1 mL) and the mixture was stirred at25° C. for 0.5 hr. The reaction mixture was diluted with H₂O (10 mL) andextracted with EtOAc (10 mL×2). The combined organic layers were driedover Na₂SO₄, filtered and concentrated. The residue was purified byprep-TLC (SiO₂, Petroleum ether:Ethyl acetate=1:1) to givetrans-isopropyl N-[4-[5-[4-(benzylcarbamoylamino)-2-[[2-[tert-butyl(dimethyl)silyl]oxy-1,1-dimethyl-ethyl]sulfamoyl]phenyl]thiazol-2-yl]cyclohexyl]carbamate(93 mg, crude) as a yellow gum. ESI [M+H]=758.4

Preparation of Ex. 74.

A mixture of trans-isopropylN-[4-[5-[4-(benzylcarbamoylamino)-2-[[2-[tert-butyl(dimethyl)silyl]oxy-1,1-dimethyl-ethyl]sulfamoyl]phenyl]thiazol-2-yl]cyclohexyl]carbamate(84.9 mg, 112 umol, 1 eq.) in AcOH (0.5 mL)/THF (0.5 mL)/H₂O (0.5 mL)was stirred at 80° C. for 0.5 hr. Then the mixture was concentrated andthe residue was purified by prep-HPLC (TFA condition) to givetrans-isopropylN-[4-[5-[4-(benzylcarbamoylamino)-2-[(2-hydroxy-1,1-dimethyl-ethyl)sulfamoyl]phenyl]thiazol-2-yl]cyclohexyl]carbamate(5.86 mg, 9.10 umol, 8.13% yield, 100% purity) as a yellow solid. ¹H NMR(400 MHz, METHANOL-d4) δ=8.25 (d, J=2.6 Hz, 1H), 7.81-7.66 (m, 2H),7.40-7.30 (m, 5H), 7.28-7.22 (m, 1H), 4.82 (td, J=6.0, 12.5 Hz, 1H),4.41 (s, 2H), 3.45 (br t, J=11.6 Hz, 1H), 3.28 (s, 2H), 3.00 (br t,J=11.8 Hz, 1H), 2.23 (br d, J=13.2 Hz, 2H), 2.11-2.00 (m, 2H), 1.77-1.60(m, 2H), 1.48-1.32 (m, 2H), 1.22 (br d, J=6.1 Hz, 6H), 1.05 (s, 6H). ESI[M+H]=644.3

Example 75 Compound Primary Screening

1. Background

Primary screening was a phenotypic screen that utilized the syntheticlethal interaction between AID and RAD51 to identify compounds that wereboth potent and on target. AID expressing cells are dependent upon RAD51for survival; inhibiting RAD51 in AID positive cells results in acytotoxic effect. Based on such an effect, compounds that were potent inAID positive cells and were significantly less potent in AID negativecells were identified.

2. Materials and Supplies

Plastic ware and consumables needed for this experiment include: CellCulture media; Evaporation Buffer media; 100% DMSO; 96 well U-bottomsterile culture plates; 250 mL bottle; 1.5 mL Opaque amber epi tubes;Epi Tube rack; 300 mL reservoirs; 25 mL reservoir; 25 mL serologicalpipette tips; 5 mL serological pipette tips P1000 Pipette Tips; and P200Pipette Tips.

Equipment needed for this experiment include: Viaflo 384 liquid handler;Eppendorf serological pipette; Eppendorf P1000 Pipette; and EppendorfP200 Pipette

Daudi Cell Culture and WI-38 Cell Cultures werealso needed for thisexperiment.

Lastly, compounds (e.g., the compounds of this invention) to be testedare needed.

3. Procedure

All steps were performed in a sterile environment inside the Biosafetycabinet.

The first step was to set up a cell killing assay in the Daudi cell line(AID positive). A 96 well u-bottom plate was prepared by writing theexperiment number, plate number, date and initials in the top rightcorner of the plate lid. With a sterile 300 ml reservoir, and 25 mlserological pipette, evaporation buffer media was pipetted intoreservoir in 25 ml increments. Using the liquid handler, 150ul ofevaporation buffer media was pipetted from reservoir into rows A and H,and Columns 1 and 12 of the 96 well u-bottom plate. Cell cultures werecounted to obtain the density of cells per ml, and the cultureviability. The cell density information was used to obtain 1,000,000cells from culture using a 5 mL serological pipette into an epi tube.The cell density information from the culture was used to calculate thenumber of cells and volume of media needed for the assay to seed 1250cells in 130ul of media per available culture well in the 96 wellu-bottom plate. Rows B through F were used for cells (50 wells intotal), with row G left for an empty media control. The calculation wasoverestimated by 10 mL to account for the dead volume in the 300 mlreservoir. Once the media volume was calculated, the appropriate volumeof media was pipetted in 25 mL increments into the 250 mL bottle using a25 mL serological pipette. The 250 ml bottle was capped tightly, andplaced into a 37° C. water bath for 2 minutes. While the culture mediawas warming, 10 mL of fresh media was pipetted from the 500 mL culturemedia bottle into a sterile 25 mL reservoir. Using the Eppendorfmultichannel pipette, 130ul of media was piptted from the 25 mLreservoir into row G of the 96 well u-bottom plate. Once the 250 mLbottle of media was warmed, the volume of culture needed was pipettedinto the bottle, and mixed gently with a 25 mL serological pipette as tonot create bubbles, and then the contents of the bottle were pipettedinto a new 300 mL reservoir. Using the liquid handler, 130ul of culturewas pipetted from the 300 mL reservoir into rows B through F of the 96well u-bottom plate. Once the culture was added, the plate was placedinto a 37° C. incubator until the compound master plate was prepared foruse.

Two 96 well u-bottom plates were prepared by writing the master platename in the upper right corner of the plate lid. Labeling one DMSOmaster and the other Media Master. The compounds of interest wereobtained from the laboratory freezer, and placed into a 25 well storagebox with a lid, and set the box aside. The compounds were vortexed afterthawing but before use. Using an automatic multichannel pipette, 20ul of100% DMSO was pipetted into wells B3-B11 through G3-G11 of the DMSOmaster plate. For each compound on the master plate, 50ul of thecompound were pipetted in the appropriate well of row 2 (reference platemap to determine appropriate well). A serial dilution was preparedbeginning by aspirating 20ul from row 2 and mixing with row 3, repeatinguntil row 11 was reached. Using the liquid handler, 194ul of Daudi mediawas dispensed into wells B2-B 11 through G2-G11 of the Media masterplate. Using the liquid handler, 6ul from the DMSO master plate wasaspirated and dispensed into the media master plate, mixing 100ul twice.

Compounds from master plate were then added to the culture plate. Theculture plates were removed from the incubator, and set inside thebiosafety cabinet. Using a liquid handler, 20ul from wells B2 to B11through G2 to G11 of master plate were aspirated, and dispensed intowells B2 to B11 through G2 to G11 of culture plate. This set wascontinued with each culture plate. Once the culture plates acquiredtheir 20ul of compound dilutions, they were placed back into theincubator, until their reads on Day 7 of experiment. Cell death wasmeasured on Day 7 of the experiment using Cell-Titer Glo and a PromegaPlate reader.

Percent cell death and EC₅₀ values were calculated by comparing the cellviability of the compound treated wells to the non-treated wells.Normalized RLU values were obtained by subtracting the media well valuesfrom each of the wells in the same column, and then dividing that valueby the DMSO treated cells values. The percent kill was then calculatedby subtracting the normalized RLU value from 1 and multiplying by 100.The average normalized percent kill value and standard error of the meanwas then calculated. The kill values were then inputted into Prism withthe corresponding standard errors. In Prism a non-linear regression linewas plotted with the data points using a semi-log scale, and the EC₅₀value was calculated. For compounds that showed good potency in theDaudi cell line, the assay was repeated using WI-38 cells (AIDnegative).

Screening Data

TABLE 1 AID + EC₅₀ AID-EC₅₀ (μM) (μM) A = ≤0.1 μM A = ≤0.1 μM B = ≤1 μMB = ≤1 μM C = >1 μM C = >1 μM Compound ND = Not ND = Not No. StructureDetermined Determined Ex. 1

C ND Ex. 2

C ND Ex. 3

A C Ex. 4

C C Ex. 5

C ND Ex. 6

C ND Ex. 7

B ND Ex. 8

C ND Ex. 9

B ND Ex. 10

A C Ex. 11

B ND Ex. 12

B ND Ex. 13

A C Ex. 14

A C Ex. 15

A C Ex. 16

A C Ex. 17

B C Ex. 18

A C Ex. 19

A C Ex. 20

C ND Ex. 21

A ND Ex. 22

B ND Ex. 23

A ND Ex. 24

A C Ex. 25

A ND Ex. 26

A C Ex. 27

A C Ex. 28

A ND Ex. 29

A C Ex. 30

A C Ex. 31

A C Ex. 32

A C Ex. 33

C ND Ex. 34

B ND Ex. 35

B ND Ex. 36

B ND Ex. 37

B ND Ex. 38

A C Ex. 39

A C Ex. 40

A C Ex. 41

A C Ex. 42

A C Ex. 43

B C Ex. 44

C ND Ex. 45

A ND Ex. 46

C C Ex. 47

A C Ex. 48

B C Ex. 49

B C Ex. 50

A C Ex. 51

A C Ex. 52

A C Ex. 53

A C Ex. 54

A C Ex. 55

A C Ex. 56

B ND Ex. 57

A C Ex. 58

B C Ex. 59

A C Ex. 59A

B C Ex. 59B

A C Ex. 60

B C Ex. 60A

C C Ex. 60B

B C Ex. 61

C C Ex. 62

C C Ex. 63

B ND Ex. 64

A C Ex. 65

B C Ex. 66A

A C Ex. 66B

B ND Ex. 67A

B C Ex. 67B

C ND Ex. 68

B ND Ex. 69

C ND Ex. 70

A C Ex. 71

A C Ex. 72

A C Ex. 73

A C Ex. 74

A C

Example 76. Bi-Directional Caco-2 Permeability

Bi-directional Caco-2 permeability was assayed. Caco-2 cells were seededonto permeable polycarbonate supports and allowed to differentiate forabout 3 weeks prior to being used in the assays. The cells were thenexposed to the compounds from either the apical or basolateral sides andincubated at 37 C for up to 90 minutes under light agitation. Compoundtransport was then measured using LC/MS/MS analysis at 30, 60, and 90minutes.

TABLE 2 Caco-2 Results AB Papp BA Papp Exam- (cm/sec × (cm/sec × BA/ABAB BA ple No. 10⁶) 10⁶) Ratio Recovery % Recovery % Ex. 5 5.9 2.2 0.434.7 80.8 Ex. 7 1.7 2.1 1.2 55.9 92 Ex. 10 1.2 1.5 1.3 59.9 87.2 Ex. 172.8 2.1 0.8 47.2 81.9 Ex. 20 9.4 10.4 1.1 44.8 96.6 Ex. 21 3.6 10.7 358.6 78.3 Ex. 22 1.1 21 18.7 83.1 89 Ex. 27 0.7 0.5 0.7 92 107.1 Ex. 281.9 3.3 1.7 66.9 81 Ex. 29 10.9 24.1 2.2 93.9 90.9 Ex. 30 11.5 15.7 1.479.8 115.6 Ex. 31 12.8 13.6 1.1 70.5 92.3 Ex. 34 0.4 45.2 103.9 98.296.8 Ex. 35 0.4 41 99.1 98.8 107.5 Ex. 38 17.9 22.9 1.3 73.3 82.3 Ex. 443.2 4.2 1.3 36.2 69.4 Ex. 45 7.6 10.8 1.4 73.6 84.4 Ex. 47 8.7 12.4 1.465.3 80.7 Ex. 48 23.1 16 0.7 80.1 90.3 Ex. 50 2 29.1 14.6 87.7 98.1 Ex.51 9.9 10.5 1.1 65 85.7 Ex. 52 7.4 10.9 1.5 64.1 91.7 Ex. 53 9.3 9.7 163.1 90.9 Ex. 55 6.5 7 1.1 65.2 83.8 Ex. 57 1.3 3.5 2.7 61.9 86.8 Ex. 585.1 3.4 0.7 52.3 83.5 Ex. 59 3.1 15 4.8 61.5 91.5 Ex. 59A 3.1 9.6 3.155.6 85.5 Ex. 59B 4.6 9.7 2.1 56.2 82 Ex. 60 15.6 13.3 0.9 59.8 90.8 Ex.60A 12 11.2 0.9 55.3 86 Ex. 60B 11.6 13.5 1.2 51.2 84.3 Ex. 63 10.5 26.72.5 83.6 90 Ex. 64 2.2 27.4 12.5 79.7 95.9 Ex. 66A 2.9 3.9 1.4 64.9 81.2Ex. 67A 10.5 6.9 0.7 50.7 78.6

Example 77. Human Liver Microsome Stability

The stability of the claimed compounds was determined in the presencesof human liver microsomes. The compounds were incubated with themicrosomes at 37° C. for 45 minutes. Samples were analyzed usingLC-MS/MS. Data analysis included half-life, clearance rate, and thepercentage of hepatic blood flow (% QH) for each of the compounds in thedifferent species. Below are liver microsome assat data ofrepresentative compounds, which show that the claimed compounds havesuperior metabolic stability.

TABLE 3 Human Liver Microsome Stability Half Life Clearance (min)(μg/min/mg) % QH Ex. 29 20.5 68.0 78.8 Ex. 31 22.4 61.9 77.3 Ex. 66A77.6 18.2 49.6 Ex. 67A >300 <4.6 <20.3

Example 78. Cell Line Screen

The activity of the claimed compounds was measured in a variety of celllines with different expression levels of activation induced cytidinedeaminase (AICDA). The potency assay was repeated in all of the listedcell lines and the EC₅₀ values recorded.

TABLE 4 EC₅₀ EC₅₀ EC₅₀ EC₅₀ Cell Line AICDA (nM) (nM) (nM) (nM) (CancerType) Expression Ex. 29 Ex. 31 Ex. 66A Ex. 67A Daudi High 43 20 18 311(Lymphoma) WSU-FSCCL Negative 67 <40 25 344 (Lymphoma) U-698-M High 11331 88 791 (Lymphoma) CCRF-SB High 1283 2164 183 932 (Leukemia) KYSE-70Low 4660 4701 2639 2629 (Head and Neck) SNU-1 Negative n.d. n.d. 6092927 (Gastric) KG-1 Negative >10000 8785 3067 2995 (Leukemia) KYSE-510Negative >10000 >10000 4516 3403 (Head and Neck) SNU-5 Low n.d. n.d.2941 3845 (Gastric) TOV-1120D Negative 9172 n.d. 2377 4924 (Ovary) OV56Low 9086 n.d. 5944 7228 (Ovary) ARPE19/HPV16Negative >10000 >10000 >10000 >10000 (HPV Immortalized RPE) WI-38Negative >10000 >10000 >10000 >10000 (Normal Human Lung Fibroblast) n.d.not determined

Example 79. Pharmacokinetic (PK)

PK studies in mice were used to determine the fate of the compounds in awhole organism. Rats were treated with the compounds either orally ofvia IV at the indicated doses and followed for up to 24 hours. Plasmasamples were taken at different time points and analyzed by LC-MS.

TABLE 5 po @ 80 mg/kg (Formulation: 30% PEG400, 10% Vitamin E TPGS inwater) Ex. 29 Ex. 31 Ex. 66A Ex. 67A Rat Female T_(1/2) (hr) 4.66 4.752.59 11.5 F (%) @ 5 mg/kg 8.39 2.77 3.31 86.5 Rat Male T_(1/2) (hr) 3.973.79 1.86 6.46 F (%) @ 5 mg/kg 3.69 1.49 2.55 46.9

What is claimed is:
 1. A compound represented by the followingstructural formula:

or a pharmaceutically acceptable salt thereof, wherein: the thiazolering is optionally substituted with F or Cl; Cy is (C₃-C₇)cycloalkyl,bridged (C₆-C₁₂)cycloalkyl, or a 4-10 membered heterocyclic ring, eachof which is optionally substituted with one or more groups selected fromthe group consisting of halogen, OH, (C₁-C₄)alkyl, and (C₁-C₄)alkoxy;when X⁵ is connected with a nitrogen ring atom of Cy, X⁵ is absent; whenX⁵ is connected with a carbon ring atom of Cy, X⁵ is NR^(a) or O; X⁶ isNR^(a) or O; R¹ is (C₁-C₅)alkyl optionally substituted with OH; R³ is(C₁-C₅)alkyl, —CH₂-phenyl, (C₃-C₇)cycloalkyl, —CH₂—(C₃-C₇)cycloalkyl,—CH₂-monocyclic 3-7 membered heterocyclic ring, or monocyclic 3-7membered heterocyclic ring, wherein the (C₁-C₅)alkyl, (C₃-C₇)cycloalkyl,phenyl, or monocyclic 3-7 membered heterocyclic ring represented by R³or in the group represented by R³ is optionally substituted with one ormore groups selected from the group consisting of halogen, OH,(C₁-C₄)alkyl, halomethyl, halomethoxy, CN, and (C₁-C₄)alkoxy; R² is—NR^(a)C(O)O(C₁-C₄)alkyl, —NR^(a)C(O)NR^(a)(C₁-C₄)alkyl,—NR^(a)C(O)O(C₂-C₄)alkenyl, —NR^(a)C(O)NR^(a)(C₂-C₄)alkenyl,—NR^(a)C(O)O—(C₃-C₆)cycloalkyl, —NR^(a)C(O)NR^(a)—(C₃-C₇)cycloalkyl,—NR^(a)C(O)O-phenyl, —NR^(a)C(O)NR^(a)-phenyl, —NR^(a)C(O)O-monocyclic3-7 membered heterocyclic ring, —NR^(a)C(O)NR^(a)-monocyclic 3-7membered heterocyclic ring, —NR^(a)C(O)O-monocyclic 5-6 memberedheteroaromatic ring, or —NR^(a)C(O)NR^(a)-monocyclic 5-6 memberedheteroaromatic ring; wherein the (C₁-C₄)alkyl and the (C₂-C₄)alkenyl inthe group represented by R² are each optionally and independentlysubstituted with one or more groups selected from the group consistingof halogen, N₃, OR^(a), NR^(a)R^(a), (C₃-C₆)cycloalkyl, phenyl, amonocyclic 3-7 membered heterocyclic ring, and a monocyclic 5-6 memberedheteroaromatic ring; wherein the (C₃-C₇)cycloalkyl in the grouprepresented by R² is optionally substituted with one or more groupsselected from the group consisting of halogen, CH₃, =O, OR^(a), andNR^(a)R^(a); wherein the phenyl in the group represented by R² isoptionally substituted with one or more groups selected from the groupconsisting of halogen, CH₃, halomethyl, halomethoxy, CN, OR^(a), and N₃;wherein the heterocyclic ring in the group represented by R² isoptionally substituted with one or more groups selected from the groupconsisting of ═O, halogen, OR^(a), CH₃, halomethyl, and halomethoxy;wherein the heteroaromatic ring in the group represented by R² isoptionally substituted with one or more groups selected from the groupconsisting of halogen, CN, CH₃, halomethyl, halomethoxy, OR^(a), andNR^(a)R^(a); and each R^(a) is independently H or CH₃.
 2. The compoundof claim 1, represented by the following structural formula:

or a pharmaceutically acceptable salt thereof, wherein: the thiazolering is optionally substituted with F or Cl; Cy is cyclohexyl or a6-membered monocyclic heterocyclic ring; X⁵ and X⁶ are eachindependently NR^(a) or O; R¹ is (C₁-C₅)alkyl; R³ is (C₁-C₅)alkyl ormonocyclic 3-7-membered heterocyclic ring; R² is—NR^(a)C(O)O(C₁-C₄)alkyl, —NR^(a)C(O)NR^(a)(C₁-C₄)alkyl,—NR^(a)C(O)O(C₂-C₄)alkenyl, —NR^(a)C(O)NR^(a)(C₂-C₄)alkenyl,—NR^(a)C(O)—O(C₃-C₆)cycloalkyl, —NR^(a)C(O)NR^(a)—(C₃-C₆)cycloalkyl,—NR^(a)C(O)O-phenyl, —NR^(a)C(O)NR^(a)-phenyl, —NR^(a)C(O)O-monocyclic3-7 membered heterocyclic ring, —NR^(a)C(O)NR^(a)-monocyclic 3-7membered heterocyclic ring, —NR^(a)C(O)O-monocyclic 5-6 memberedheteroaromatic ring, or —NR^(a)C(O)NR^(a)-monocyclic 5-6 memberedheteroaromatic ring; wherein the (C₁-C₄)alkyl and the (C₂-C₄)alkenyl inthe group represented by R² are each optionally and independentlysubstituted with one or more halogen, N₃, OR^(a), NR^(a)R^(a),(C₃-C₆)cycloalkyl, phenyl, monocyclic 3-7-membered heterocyclic ring, ormonocyclic 5-6-membered heteroaromatic ring; wherein the(C₃-C₆)cycloalkyl in the group represented by R² is optionallysubstituted with one or more halogen, CH₃, OR^(a), or NR^(a)R^(a);wherein the phenyl in the group represented by R² is optionallysubstituted with one or more halogen, CH₃, halomethyl, halomethoxy,OR^(a) or N₃; wherein the heterocyclic ring in the group represented byR² is optionally substituted with one or more ═O, halogen, CH₃,halomethyl, or halomethoxy; wherein the heteroaromatic ring in the grouprepresented by R² is optionally substituted with one or more halogen,CH₃, halomethyl, halomethoxy, OR^(a), or NR^(a)R^(a); and each R^(a) isindependently H or CH₃.
 3. The compound of claim 1 or a pharmaceuticallyacceptable salt thereof, wherein Cy is cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, azetidinyl, azepanyl,diazaspiro[4.4]nonyl, diazaspiro[3.5]nonyl, diazepanyl,dihydroimidazolyl, dihydrofuranyl, dihydropyranyl, dihydropyridinyl,dihydropyrimidinyl, dihydrothienyl, dihydrothiophenyl,dihydrothiopyranyl, hexahydropyridazinyl, hexahydropyrimidinyl,hydantoinyl, indolinyl, isoindolinyl, morpholinyl, oxiranyl, oxetanyl,piperidinyl, piperazinyl, pyrrolidinonyl, pyrrolidinyl,tetrahydrofuranyl, tetrahydroimidazolyl, tetrahydroindolyl,tetrahydropyranyl, tetrahydrothienyl, tetrahydropyridinyl,tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl,thiomorpholinyl, tropanyl, valerolactamyl, bicyclo[2.1.1]hexyl,bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, bicyclo[3.2.1]octyl,bicyclo[4.3.1]decyl, bicyclo[3.3.1]nonyl, bornyl, bornenyl, norbornyl,norbornenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, tricyclobutyl, adamantly,azanorbornyl, quinuclidinyl, isoquinuclidinyl, tropanyl,azabicyclo[2.2.1]heptanyl, 2-azabicyclo[3.2.1]octanyl,azabicyclo[3.2.1]octanyl, azabicyclo[3.2.2]nonanyl,azabicyclo[3.3.0]nonanyl, azabicyclo [3.3.1]nonanyl,diazabicyclo[2.2.1]heptanyl, diazabicyclo[3.2.1]octanyl,octahydropyrrolo[3,4-b]pyrrolyl, or octahydropyrrolo[3,4-c]pyrrolyl. 4.The compound of claim 1, represented by the following structuralformula:

or a pharmaceutically acceptable salt thereof, wherein: X⁷ is NH or O;R⁴ is (C₁-C₄)alkyl, (C₃-C₆)cycloalkyl, or a monocyclic 3-7 memberedheterocyclic ring; wherein the (C₁-C₄)alkyl represented by R⁴ isoptionally substituted with one or more groups selected from the groupconsisting of halogen, N₃, OR^(a), NR^(a)R^(a) (C₃-C₆)cycloalkyl,phenyl, a monocyclic 3-7 membered heterocyclic ring, and a monocyclic5-6 membered heteroaromatic ring, wherein the (C₃-C₆)cycloalkyl or themonocyclic 3-7 membered heterocyclic ring represented by R⁴ or the(C₃-C₆)cycloalkyl or the monocyclic 3-7 membered heterocyclic ring inthe group represented by R⁴ is optionally substituted with one or moregroups selected from the group consisting of halogen, OR^(a), ═O, andCH₃, wherein the phenyl in the group represented by R⁴ is optionallysubstituted with one or more groups selected from the group consistingof halogen, CH₃, halomethyl, halomethoxy, OR^(a), and N₃; wherein theheteroaromatic ring in the group represented by R⁴ is optionallysubstituted with one or more groups selected from the group consistingof halogen and CH₃.
 5. The compound of claim 4 or a pharmaceuticallyacceptable salt thereof, wherein: X⁷ is NH or O; R³ is (C₁-C₅)alkyl; R⁴is (C₁-C₄)alkyl optionally substituted with one or more halogen, OR^(a),NR^(a)R^(a), (C₃-C₆)cycloalkyl, phenyl, monocyclic 3-7-memberedheterocyclic ring, or monocyclic 5-6-membered heteroaromatic ring;wherein the phenyl is optionally substituted with one or more halogen,CH₃, halomethyl, halomethoxy, OR^(a), or N₃; wherein the 3-7-memberedheterocyclic ring is optionally substituted with one or more ═O,halogen, or CH₃; and wherein the 5-6-membered heteroaromatic ring isoptionally substituted with one or more halogen or CH₃.
 6. The compoundof claim 1, selected from the following structural formulae:

or a pharmaceutically acceptable salt thereof.
 7. The compound of claim1 or a pharmaceutically acceptable salt thereof, wherein Cy isazetidinyl or pyrrolidinyl, and the nitrogen ring atom is connected withthe thiazole ring.
 8. The compound of claim 1 or a pharmaceuticallyacceptable salt thereof, wherein Cy is 1,7-diazaspiro[4.4]nonyl,2,7-diazaspiro[4.4]nonyl, 2,7-diazaspiro[3.5]nonyl, 1,4-diazepanyl,2,5-diazabicyclo[2.2.1]heptanyl, 3,8-diazabicyclo[3.2.1]octanyl,octahydropyrrolo[3,4-b]pyrrolyl, or octahydropyrrolo[3,4-c]pyrrolyl, andthe two nitrogen ring atoms are connected with the thiazole ring and theX⁵C(O)X⁶R³ moiety, respectively.
 9. The compound of claim 4 or apharmaceutically acceptable salt thereof, wherein R⁴ is (C₁-C₃)alkyl,(C₃-C₆)cycloalkyl, or a monocyclic 3-7 membered heterocyclic ring,wherein the (C₁-C₃)alkyl is optionally substituted with (i) phenyloptionally substituted by one or more halogen or CH₃; (ii) a monocyclic5-6 membered heteroaromatic ring optionally substituted by one or morehalogen or CH₃; or (iii) a monocyclic 3-7 membered heterocyclic ringoptionally substituted by one or more halogen or CH₃.
 10. The compoundof claim 4 or a pharmaceutically acceptable salt thereof, wherein R⁴ is(C₁-C₃)alkyl, CH₂-phenyl, CH₂-5-6 membered heteroaromatic ring, orCH₂-3-7 membered monocyclic heterocyclic ring, wherein the phenyl, 5-6membered heteroaromatic ring, or 3-7 membered monocyclic heterocyclicring in the group represented by R⁴ is optionally substituted by one ormore groups selected from the group consisting of halogen and CH₃. 11.The compound of claim 4 or a pharmaceutically acceptable salt thereof,wherein R⁴ is (C₁-C₃)alkyl optionally substituted with (i) phenyloptionally substituted by one or more halogen, CH₃, halomethyl,halomethoxy, OR^(a), or N₃; (ii) a monocyclic 5-6-memberedheteroaromatic ring optionally substituted by one or more halogen orCH₃; or (iii) a monocyclic 3-7_membered heterocyclic ring optionallysubstituted by one or more ═O or CH₃.
 12. The compound of claim 4 or apharmaceutically acceptable salt thereof, wherein R⁴ is (i)(C₁-C₃)alkyl; (ii) CH₂-phenyl optionally substituted by halogen, CH₃,halomethyl, halomethoxy, OR^(a), or N₃; (iii) CH(CH₃)-phenyl optionallysubstituted by halogen, CH₃, halomethyl, halomethoxy, OR^(a), or N₃;(iv) CH₂-5-6 membered heteroaromatic ring optionally substituted byhalogen or CH₃; or (v) CH₂-3-7_membered monocyclic heterocyclic ringoptionally substituted by ═O or CH₃.
 13. The compound of claim 1 or apharmaceutically acceptable salt thereof, wherein R³ is (C₁-C₄)alkyl,(C₄-C₆)cycloalkyl, CH₂-phenyl, CH₂-monocyclic 4-6 membered heterocyclicring, or monocyclic 4-6 membered heterocyclic ring, wherein the phenylor monocyclic 4-6 membered heterocyclic ring represented by R³ or in thegroup represented by R³ is optionally substituted with one or moregroups selected from the group consisting of halogen, OH, OCH₃, and CH₃.14. The compound of claim 1, selected from the following structuralformulae:

or a pharmaceutically acceptable salt thereof.
 15. The compound of claim1 or a pharmaceutically acceptable salt thereof, wherein R³ isisopropyl, tert-butyl, cyclobutyl, cyclopentyl, benzyl, oxetanyl,tetrahydro-2H-pyranyl, or


16. The compound of claim 1 or a pharmaceutically acceptable saltthereof, wherein R¹ is tert-butyl.
 17. The compound of claim 1 or apharmaceutically acceptable salt thereof, wherein R³ is isopropyl oroxetanyl.
 18. The compound of claim 1 or a pharmaceutically acceptablesalt thereof, wherein R⁴ is


19. A pharmaceutical composition comprising a pharmaceuticallyacceptable carrier or diluent and a compound of claim 1 or apharmaceutically acceptable salt thereof.